JP2020517735A - Composition comprising Propionibacterium acnes bacteriophage for treating acne - Google Patents

Abstract

In particular, provided herein are compositions, systems and methods for preventing or treating acne. Compositions, combinations, systems and methods comprising at least one Propionibacterium acnes bacteriophage, at least one anti-acne compound and a pharmaceutically acceptable carrier are included. Also included are compositions, combinations and systems comprising Propionibacterium acnes bacteriophage and enzymes. Methods for preventing or treating acne are also provided.

Description

This application claims the benefit of priority of US Provisional Application No. 62/488,326, filed April 21, 2017, which is hereby incorporated by reference in its entirety for all purposes. Incorporate.

This invention was made with government support under Grant No. 1R43AR0687172-01 awarded by the National Institutes of Health. The government has certain rights in this invention.

The content of the text file named “052004-503001 WO_Sequence List.TXT” is created on April 20, 2018 and has a size of 101,782 bytes, which is incorporated by reference in its entirety.

Acne is a nearly universal condition affecting more than 80% of all people in the world. Although this chronic skin condition is complex, the major pathogen is Propionibacterium acnes, whose overgrowth leads to comedone-induced inflammation. Despite the clear need for innovation, there have been no new comedones for over 30 years. Current treatments, including benzoyl peroxide and antibiotics, are extremely ineffective, and the most effective treatment, isotretinoin, is minimal due to dangerous side effects (including birth defects, liver damage and suicide). It is limited to patients.

There is a need for new methods and compositions for treating acne.

In particular, provided herein are compositions, combinations, systems and methods for preventing or treating acne.

In an aspect, a composition herein comprising, consisting essentially of, or consisting of at least one Propionibacterium acnes bacteriophage, at least one anti-acne compound and a pharmaceutically acceptable carrier. Provided by.

In an aspect, provided herein is a composition comprising at least one Propionibacterium acnes bacteriophage, one or less anti-acne compound, and a pharmaceutically acceptable carrier.

In an aspect, provided herein is a composition comprising an active ingredient consisting of at least one Propionibacterium acnes bacteriophage and one or less anti-acne compound, and a pharmaceutically acceptable carrier.

In an aspect, provided herein is a composition comprising at least one Propionibacterium acnes bacteriophage, at least one anti-acne compound and a pharmaceutically acceptable carrier, and free of probiotic bacteria.

In an aspect, provided herein is a composition comprising a Propionibacterium acnes bacteriophage and an enzyme.

In an aspect, a combination comprising, consisting essentially of, or consisting of at least one Propionibacterium acnes bacteriophage and at least one anti-acne compound, wherein the at least one Propionibacterium acnes Provided herein are combinations of bacteriophage and at least one anti-acne compound, each in a composition further comprising a pharmaceutically acceptable carrier.

In an aspect, provided herein is a combination comprising a Propionibacterium acnes bacteriophage and an enzyme.

In an aspect, a method of preventing or treating acne in a subject in need thereof is provided by administering an effective amount of a composition or combination provided herein. Provided herein.

P. Acnes (causing acne, left half plate) or P. P. granulosum (symbiotic, right half plate) bacteria were plated on RCM agar Petri dishes. Sterile half-pads dipped in either minocycline or PHIT-101 (10 ⁷ pfu/mL) were placed on each plate. After anaerobic incubation at 37°C for 3 days, a germicidal zone (arrow) appears, minocycline kills both pathogens and commensal bacteria, and PHIT-101 kills acne-causing bacteria and symbiotic P. It has been shown to not harm Granulosa. P. Acnes, P. Granulosa and P. A synthetic skin microbiome containing P. avidum was grown to confluence in vitro. The synthetic skin microbiome was then incubated for 48 hours in the presence or absence of PHIT-101. The composition ratio of the three bacterial species was quantified by NGS sequencing of the 16S amplification product of the bacterial pellet washed using the Illumina MiSeq platform. PHIT-101 is a P. Acnes could be almost completely annihilated and had no effect on the growth of the other two symbiotic species. P. Biofilm production within Acnes is highly variable. 96 strains of P. Acnes was grown in 96-well polystyrene microtiter plates to stimulate biofilm production and the biofilm produced by each strain was quantified. The variability demonstrated within this set of strains demonstrates the need to quantify biofilm formation under growth conditions more similar to those found in human pores. P. Screening to select enzymes capable of degrading Acnes biofilm. P. Acnes was grown in polystyrene microtiter plates to stimulate biofilm production. Enzyme was added to wells at 0.01 mg/mL and incubated at 30°C for 30 minutes. Degraded biofilm was washed away with phosphate buffered saline (PBS) and residual biofilm in each well was quantified by staining with crystal violet and recording the absorbance at 590 nm. Proteases such as proteinase K and subtilisin showed good activity, and dispersin was the best glycoside depolymerase tested. Augmentation of phage with biofilm degrading enzyme (BDE) greatly increases bactericidal capacity. Sticking P. Acnes cells were incubated with PBS (untreated), PHIT-101, or PHIT-101 and dispersin. Cell viability was measured using CellTiter-Blue reagent and fluorescence was recorded at 560 _Ex /590 _Em . PHIT-101 is more effective than P. Although Acnes could not be killed, the addition of the biofilm-degrading enzyme Dispersin significantly increased the killing capacity to a level similar to liquid culture. Probiotic strains produce low levels of lipase in adherent culture. Probiotic P. cerevisiae with a known genotype. The Acnes strain was grown under biofilm conditions in microtiter plates. After 72 hours of growth, the culture supernatant was filter-sterilized and incubated with 4-MU palmitate for 4 hours at 37° C. to determine extracellular lipase production. The lipase production of the probiotic strain (Pr#X) is much lower than that of the pathogen, indicating relatively low proinflammatory properties. Probiotic strains are pathogenic P. Significantly less adherence to epithelial cells than Acnes. Selected probiotic strains were incubated with confluent A-431 epithelial cells (MOI10). After washing the wells, cells were lifted using 0.1% Tween 80 and seeded on BHI plates. Colonies were counted after 72 hours of anaerobic incubation. The data revealed that the probiotic strain showed significantly lower binding to epithelial cells ( ^* p<0.05, ^** p<0.005). Lower pro-inflammatory properties of probiotic strains in a mouse ear inflammation model. CBA/J mice (5 mice per cohort) received P. The Acnes strain was injected and cytokine analysis was performed on day 5. The probiotic strain Pr#C had significantly lower levels ( ^* p<0.05, ^** p<0.01, ^*** p<0.0001) of the inflammatory cytokine IL- compared to the pathogenic strain. 1β (FIG. 8A), IL-6 (FIG. 8B), IL-17 (FIG. 8C) and TNFα (FIG. 8D) are shown. Pr-C has a ProII 16S sequence. Lower pro-inflammatory properties of probiotic strains in a mouse ear inflammation model. CBA/J mice (5 mice per cohort) received P. The Acnes strain was injected and cytokine analysis was performed on day 5. The probiotic strain Pr#C had significantly lower levels ( ^* p<0.05, ^** p<0.01, ^*** p<0.0001) of the inflammatory cytokine IL- compared to the pathogenic strain. 1β (FIG. 8A), IL-6 (FIG. 8B), IL-17 (FIG. 8C) and TNFα (FIG. 8D) are shown. Pr-C has a ProII 16S sequence. Lower pro-inflammatory properties of probiotic strains in a mouse ear inflammation model. CBA/J mice (5 mice per cohort) received P. The Acnes strain was injected and cytokine analysis was performed on day 5. The probiotic strain Pr#C had significantly lower levels ( ^* p<0.05, ^** p<0.01, ^*** p<0.0001) of the inflammatory cytokine IL- compared to the pathogenic strain. 1β (FIG. 8A), IL-6 (FIG. 8B), IL-17 (FIG. 8C) and TNFα (FIG. 8D) are shown. Pr-C has a ProII 16S sequence. Lower pro-inflammatory properties of probiotic strains in a mouse ear inflammation model. CBA/J mice (5 mice per cohort) received P. The Acnes strain was injected and cytokine analysis was performed on day 5. The probiotic strain Pr#C had significantly lower levels ( ^* p<0.05, ^** p<0.01, ^*** p<0.0001) of the inflammatory cytokine IL- compared to the pathogenic strain. 1β (FIG. 8A), IL-6 (FIG. 8B), IL-17 (FIG. 8C) and TNFα (FIG. 8D) are shown. Pr-C has a ProII 16S sequence. P. Acnes strains have different lipase profiles in suspension and adherent cultures. Two sets of pathogenicity (Path-1, Path-2) and two viable bacteria (Pr-1 to Pr-6) P. The Acnes strain was evaluated for lipase production in floating (grey bars) and adherent (black bars) cultures. Although lipase production of probiotic strains was not significantly different from pathogenic strains in liquid (floating) cultures, lipase production in adherent cultures was consistently lower than that in the corresponding pathogenic cultures. Interestingly, the variability of lipase production among probiotic strains was observed. The strain with the lowest lipase activity was selected. Illustrates the life cycle of an exemplary bacteriophage. Counterclockwise from bottom left: Phage particles recognize host bacteria and adsorb to their surface. The phage genome is injected into the bacterium. In the lysogen cycle, this DNA integrates into the bacterial genome and replicates with several cycles of the bacterial genome. During the lytic cycle, the phage genome is not integrated and subsequently hijacks the host machinery to replicate the phage genome and phage structural components. The fully assembled phage then lyse the cells, typically by producing endolysin and holin at a later stage of infection. At this point, the released phage are free to seek out new host bacteria and infect them, initiating another lysis cycle. Illustrates the formation of an exemplary bacterial biofilm. Bacterial cells land and adhere to surfaces that have favorable conditions for growth. Bacterial cells replicate to form colonies that eventually initiate biofilm formation at a certain threshold cell density (quorum). Biofilms contain a mixture of polysaccharides, proteins, DNA and lipids in varying proportions. Biofilms are physical barriers that protect bacterial colonies from harsh external conditions and confer resistance to antibiotics, toxins and immune cells. Collectively illustrates an embodiment of the actions of the three components; their effects are drawn sequentially for illustrative purposes. Inflamed comedones were typically overgrown with overgrown P. elegans, along with symbiotic skin bacteria (B). It is clogged with the biofilm (A) produced by Acnes. The biofilm degrading enzyme (lightning bolt) is P. Destroys acnes biofilm and improves access to other ingredients. Next, the bacteriophage (hexagon) is a pathogenic P. Edit acne or sterilize specifically to remove infection. Finally, the probiotic bacteria (C) colonize the pores and occupy the pathogen's niche, preventing the pathogen from growing and returning and reconditioning the microbiome to a healthy state. FIG. 1 is a schematic diagram of a non-limiting probiotic bacterial screening process. FIG. 6 is a graph showing that the pathogenic strain causes significantly higher ear inflammation than the PBS control, and the lead probiotic strain Pr-C induces ear inflammation that is not significantly different from the PBS control. FIG. 6 is a graph showing that phages remain stable in the presence of low (0.5% w/v) and high (2% w/v) concentrations of salicylic acid. Is a graph showing that the phage lose their viability in the presence of benzoyl peroxide for 60 days. The rate of decrease in phage viability is greater at the higher concentration (10% w/v) compared to the lower concentration (2.5% w/v).

In particular, provided herein are compositions, combinations, methods and systems for treating and preventing acne.

Salicylic acid and benzoyl peroxide are the most commonly used anti-acne agents in over-the-counter (OTC) products. In particular, the stability of phage when combined with these anti-acne agents is not known, as phage are broadly divergent in their stability and response to external physical and chemical factors. The redox nature of benzoyl peroxide and sulfur can potentially cause the degradation of phage protein membranes. Previous studies have shown that exposure to peroxide increases the rate of protein degradation by destabilizing the protein and making it more susceptible to proteolysis (Fligiel et al., Protein degradation following treatment withth. hydrogen peroxide.Am J Pathol 1984 years, 115 (3), pp. 418~25; Kocha, et al., Hydrogen peroxide-mediated degradation of protein: different oxidation modes of copper- and iron-dependent hydroxyl radicals on the degradation of albumin.Biochim Biophys Acta 1997, 1337(2), 319-26). Salicylic acid is known for its protein-binding ability (Lee et al., Protein binding of aceticylsalicylic acid and salicylic acid in porine and human serum. Vet Hum Toxicol 1995, 2-5, 37, 5; Plasma protein binding of salicylic acid, phenytoin, chloropromazine, propronol and peditheine ingulium eutriculation 6 and urticula 6), phenotochondria pneumoniae 3 urinary 6th, urinary 6th, urinary 6th, urinary 6th, urinary 6th, urinary 6th, urinary 6th, urinary 6th, urinary 6th, urinary 6th, and urinary 6th, urinary 6th, urinary 6th, and urinary 6th, and urinary 6th, and urinary 6th, and urinary 6th, and urinary 6th, respectively. It is believed to be non-viable.

Surprisingly, Propionibacterium acnes bacteriophage was found to be stable in compositions containing salicylic acid. See, for example, FIG. Thus, salicylic acid has been shown to be well tolerated by phage, and is a suitable anti-acne agent as a combination drug. In embodiments, the antikeratolytic activity of salicylic acid complements phage activity by allowing deeper penetration of the phage, thereby enhancing the bactericidal efficiency of the phage. In embodiments, the phages described herein may be combined with salicylic acid in the composition to prevent and treat acne.

Benzoyl peroxide is not suitable for combination with the tested phages (see FIG. 16), for example, in formulations that are stored for longer than a few days, while benzoyl peroxide is an anti-acne combination (eg kit It can be used with the phage product as part of In embodiments, benzoyl peroxide is the active ingredient in a cleanser and the cleanser is applied to the skin and washed off prior to application of the phage composition/formulation. In an embodiment, the antikeratolytic and transient antibacterial effects of benzoyl peroxide are demonstrated by bacteriophage P. Complements specific deeper selective killing of Acnes.

In embodiments, the Propionibacterium acnes bacteriophage and anti-acne compound (such as salicylic acid and/or sulfur) are in a single composition that is topically administered to the skin of the subject. In embodiments, a kit is provided that includes a Propionibacterium acnes bacteriophage and an anti-acne compound (eg, in separate containers such as bottles). In embodiments, the Propionibacterium acnes bacteriophage is in one composition and the anti-acne compound (such as benzoyl peroxide, salicylic acid and/or sulfur) is in another composition and each The composition is topically administered to the skin of the subject. In an embodiment, a Propionibacterium acnes bacteriophage is administered to the subject and then an anti-acne compound is administered to the subject. In an embodiment, the anti-acne compound is administered to the subject and then the Propionibacterium acnes bacteriophage is administered to the subject. In embodiments, the subject's face is washed while the anti-acne compound and the Propionibacterium acnes bacteriophage (in either order) are topically administered to the subject's face.

In embodiments, an effective amount of an anti-acne compound (such as benzoyl peroxide, salicylic acid or sulfur) when used in combination with a Propionibacterium acnes bacteriophage is when the anti-acne compound is used alone. Less than needed. In embodiments, an effective amount of an anti-acne compound (such as benzoyl peroxide, salicylic acid or sulfur) when used in combination with a Propionibacterium acnes bacteriophage is when the anti-acne compound is used alone. Less than 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 10% less than needed.

Definitions The following definitions are included for the purpose of understanding the present subject matter and for constructing the appended claims. The abbreviations used herein have their conventional meaning within the chemical and biological arts.

While various embodiments and aspects of the invention are disclosed and described herein, it will be apparent to those skilled in the art that such embodiments and aspects are provided by way of example only. Many variations, modifications and substitutions will now occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be used in carrying out the invention.

Section headings used herein are for organizational purposes only and should not be construed as limiting the subject matter described. All documents or portions of documents cited in this application include, but are not limited to, patents, patent applications, articles, books, manuals and treatises, in their entirety for any purpose. For more specific reference.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. For example, Singleton et al., DICTIONARY OF MICROBIOLOGY AND MOLECULAR BIOLOGY 2nd edition, J. Wiley & Sons (New York, NY 1994); Sambrook et al., MOLECULAR CLONING, A LABORATORY MANUAL, Cold Springs Harbor Press (Cold Springs Harbor), 1982, NY. Any methods, devices and materials similar or equivalent to those described herein can be used in the practice of the present invention. The following definitions are provided to facilitate understanding of certain terms used frequently herein and are not intended to limit the scope of the present disclosure.

As used herein, "Propionibacterium acnes bacteriophage" refers to P. It is a bacteriophage that infects Acnes cells, replicates inside them, and kills them. In an embodiment, P. Acnes bacteriophage is a lytic P. Acnes bacteriophage. In an embodiment, P. Acnes bacteriophage is P. Acnes bacteria can be lysed, and P. It cannot lyse any bacteria except Acnes. In an embodiment, P. Acnes bacteriophage cannot maintain lysogenicity in bacteria. In an embodiment, P. The use of bacteriophages that are able to lyse acnes but not lysogens does not allow the bacteriophage to remain dormant within the bacterium, but always lyses the bacterium and thus the bacterium. Has the advantage of sterilizing. In an embodiment, P. Acnes bacteriophage lacks the ability to express at least one gene required to maintain lysogenicity. The term "lacking the ability to express at least one gene necessary to maintain lysogenicity" refers to the result of one or more point mutations or complete or partial deletion of the genome. P. It is intended to show that Acnes bacteriophage lack the ability to produce the fully functional protein product required to maintain lysogenicity. In an embodiment, P. Acnes bacteriophages have a genome that lacks all or part of at least one gene required to maintain lysogenicity (eg, whether artificial or natural, eg, a strain is lysogenic). A strain lacking all or part of at least one gene required for maintenance, or derived from a strain). However, in embodiments, the P. acnes bacteriophage may affect the ability of the phage to maintain lysogenicity, such as a defect in the genome integration site (eg, a/att/site) or repressor binding site. It may also contain genomic defects (eg, mutations, insertions or deletions) in certain non-coding regions. In an embodiment, P. Acnes bacteriophage have the additional advantage of being naturally occurring and isolated, without the need for introducing artificial mutations into the bacteriophage. In an embodiment, P. Acnes bacteriophage is described in P. Multiple strains of Acnes can be lysed. In an embodiment, P. Acnes bacteriophage is described in P. At least about 5, 10, 15, 20, 25, 30 or more strains of Acnes can be lysed. P. Non-limiting examples of Acnes bacteriophage are disclosed herein. In an embodiment, P. Acnes bacteriophage has at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 95%, 99% SEQ ID NO:1. Has 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9% sequence identity. Has a genome. In an embodiment, P. The acnes bacteriophage has a genome having the sequence of SEQ ID NO:1 or comprises the sequence SEQ ID NO:1. In an embodiment, P. The Acnes bacteriophage genome has no insertions or deletions compared to SEQ ID NO:1. In an embodiment, P. The Acnes bacteriophage genome has no insertions or deletions as compared to SEQ ID NO:1 and only conservative substitutions. In an embodiment, P. Acnes bacteriophage is under the provisions of the Budapest Treaty and is deposited under the accession number under the National Collection of Industrial, Marine and Food Bacteria (NCIMB), Ferguson Building, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA, United Kingdom. P. It is one of the following exemplary isolates of Acnes bacteriophage: Accession number NCIMB 41332 (Isolate PA6); Accession number NCIMB 41334 (Isolate 1874); Accession number NCIMB 41333 (Isolate 1878); Accession number NCIMB 41335 (Isolate 1905); Accession number NCIMB 41349 (Isolate 1894); Accession number NCIMB 41350 (Isolate 103609); Accession number NCIMB 41351 (Isolate 103672). In an embodiment, the host bacterium, P. A non-limiting example of Acnes, AT1 has been deposited as NCIMB 41336. In an embodiment, P. Acnes bacteriophage has at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% in the genome of the bacteriophage deposited under accession number NCIMB 41349. It has a genome with 90%, 95% or 99% sequence identity. In an embodiment, P. Acnes bacteriophage has a genome with at least 87% sequence identity to the genome of the bacteriophage deposited under accession number NCIMB 41350. In an embodiment, P. Acnes bacteriophage has a genome with at least 88% sequence identity to the genome of the bacteriophage deposited under accession number NCIMB 41351. P. Additional non-limiting descriptions relating to Acnes bacteriophage are provided in US Pat. No. 9,068,159 B2, issued June 30, 2015, the entire contents of which are hereby incorporated by reference. Incorporate in the specification. The terms "phage" and "bacteriophage" are used interchangeably herein.

As used herein, "degrading" a biofilm means cleaving (eg, by enzymatic activity) a covalent bond of at least one compound that forms part of the biofilm. Non-limiting examples of compounds that may form part of the biofilm include polymers, glycosides, proteins, polysaccharides and nucleic acids. As used herein, "P. acnes biofilm degrading enzyme" refers to P. acnes. An enzyme that degrades at least one compound that forms part of the Acnes biofilm.

The enzymes provided herein maintain enzymatic activity (at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% active compared to the native protein. Up to) any naturally occurring form, homologue, asoform or variant. In embodiments, the variant is at least 90%, 95%, 96% over the entire sequence or a portion of the sequence (eg, 50, 100, 150 or 200 contiguous amino acid moieties) as compared to the naturally occurring form. It has 97%, 98%, 99% or 100% amino acid sequence identity.

The term "isolated", when applied to a bacterium or bacteriophage, (1) includes at least some of the components with which it was originally associated (whether natural or in an experimental setting). And/or (2) manually produced using artificial culture conditions such as, but not limited to, growing on plates and/or in fermentors, A bacterium or bacteriophage that has been prepared, purified and/or manufactured. Isolated bacteria include bacteria that are cultivated, even if such cultures are not monocultures. In embodiments, the isolated bacterium is a bacterium that is cultivated as a monoculture (eg, on a plate or in a liquid culture such as a fermentor). Isolated bacteria and bacteriophage comprise at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70% of the other components with which it was originally associated. , About 80%, about 90%, about 95%, about 96%, about 97%, about 99% or more (eg, by weight). In embodiments, the isolated bacterium is greater than about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%. About 97%, about 98%, about 99% or more than about 99% (eg, by weight) pure. In embodiments, the isolated bacteriophage is greater than about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%. , About 97%, about 98%, about 99% or more than about 99% (eg, by weight) pure. In embodiments, the compositions provided herein include one or more isolated bacteriophage. In an embodiment, the composition provided herein comprises an isolated bacteriophage. In embodiments, the bacteriophage administered is an isolated bacteriophage. In embodiments, the compositions provided herein include one or more isolated bacteria. In embodiments, the compositions provided herein include isolated bacteria. In embodiments, the bacterium administered is an isolated bacterium.

A "control" sample or value is a sample that serves as a reference, usually a known reference, for comparison with a test sample. For example, a test sample is taken from a test condition, eg, in the presence of a test compound (eg, enzyme) or phage, in the absence of the test compound, phage or bacterium (negative control) or a known compound, phage or bacterium. It is possible to compare with a sample collected under known conditions such as in the presence of (positive control). A control can also represent an average value collected from several tests or results. One of ordinary skill in the art will recognize that controls can be designed for the evaluation of any number of parameters. For example, a control is designed to compare therapeutic effects based on pharmacological data (eg, half-life, biofilm or degradation of this component, or bacterial cell lysis) or therapeutic strategies (eg, comparison of side effects). It is possible to One of ordinary skill in the art can understand which controls are useful in a given situation and analyze the data based on comparison to control values. Controls also help determine the importance of the data. For example, test sample variability is not considered significant if the values of a given parameter differ significantly in the control.

"Nucleic acid" refers to nucleotides (eg, deoxyribonucleotides or ribonucleotides) and their polymers in single-, double-, or multi-stranded form or their complements. The terms “polynucleotide”, “oligonucleotide”, “oligo” or the like refer to, in their ordinary and customary sense, a linear sequence of nucleotides. Oligonucleotides are typically about 5, 6, 7, 8, 9, 10, 12, 15, 25, 30, 40, 50 or longer nucleotides up to about 100 nucleotides in length. A polynucleotide is a polymer of any length, including longer lengths, eg, 200, 300, 500, 1000, 2000, 3000, 5000, 7000, 10000, 20000, 30,000, 40,000, etc. Polynucleotides and oligonucleotides generally contain phosphodiester bonds, but in some cases, for example, phosphoramidic acid, phosphorothioates, dithiophosphoric acids or O-methyl phosphoramidite chains (Eckstein, Oligonucleotides and Analogues: A Practical Approach, Alternative backbones, including Oxford University Press); and peptide nucleic acid backbones and nucleic acid analogs that may have linkages. Other analogs of nucleic acids are described in US Pat. No. 5,235,033 and US Pat. Nucleic acids having a positive skeleton, a non-ionic skeleton and a non-ribose skeleton, including the skeleton described above. Nucleic acids containing one or more carbocyclic sugars are also included within one definition of nucleic acids. Modification of the ribose-phosphate backbone may be done for a variety of reasons, for example to increase the stability and half-life of such molecules in physiological environments or as probes on biochips. It is possible to make mixtures of naturally occurring nucleic acids and analogs, and alternatively, mixtures of different nucleic acid analogs, and mixtures of naturally occurring nucleic acids and analogs.

The term "bp" and the like, in its ordinary and customary sense, refers to the indicated number of base pairs.

"Percentage sequence identity" is determined by comparing two optimally aligned sequences in a comparison window, where a portion of the polynucleotide or polypeptide sequence comparison window is the optimal alignment of the two sequences. Additions or deletions (ie, gaps) may be included when compared to the reference sequence for (no additions or deletions). In embodiments, the percentage determines the number of positions where the same nucleobase or amino acid residue is present in both sequences and divides the number of matched positions by the total number of positions in the comparison window, resulting in 100. It is calculated by multiplying by to give the percent sequence identity.

The term "identical" or percent "identity", in the context of two or more nucleic acid or polypeptide sequences, uses a comparison window or one of the following sequence comparison algorithms, or is manual. A specified percentage of amino acid residues or nucleotides that are the same or the same when aligned for optimal match, compared over a designated region measured by alignment and visual inspection (i.e., total nucleic acid, for example). Or 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91% over a particular region of the polypeptide sequence or individual portions or domains of the nucleic acid or polypeptide, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more) with two or more sequences or subsequences. Such sequences are then said to be "substantially identical." This definition extends to the complement of the test sequence. In embodiments, identity is about 20, 50, 100, 1000, 2500, 5000, 7500, 10000, 15000, 20000, 25000, or 30000 amino acids or nucleotides in length, or at least about 20, 50, 100, 1000, 2500. 5000, 7500, 10000, 15000, 20000, 25000, or 30000 amino acids or nucleotide length to about 31000, 32000, 33000, 34000 or 35000 amino acids or nucleotide length, about 31000, 32000, 33000, 34000 or 35000 amino acids or less than nucleotide length , Or at least about 31000, 32000, 33000, 34000 or 35000 amino acids or nucleotides in length. Optionally, identity exists over a region that is at least about 10 to about 100, about 20 to about 75, about 30 to about 50 amino acids or nucleotides in length. Optionally, identity exists over a region that is at least about 50 amino acids in length, or more preferably over a region that is 100-500 or 1000 or more amino acids in length. A phage comprising a nucleic acid (eg, a genome or a portion thereof) having a sequence that is substantially identical to any of SEQ ID NOs: 1, 11, 13, 15, 17, 19, 21, 23, 25 or 27. Included herein. Non-limiting examples of phage provided herein include a genome having a sequence that is substantially identical to SEQ ID NO:1.

In sequence comparison, typically one sequence acts as a reference sequence, to which test sequences are compared. When using a sequence comparison algorithm, test and reference sequences are placed in a computer, subsequence coordinates are designated, if necessary, and sequence algorithm program parameters are designated. Preferably, default program parameters can be used, or alternative parameters can be designated. The sequence comparison algorithm then calculates the percent sequence identities for the test sequences relative to the reference sequence, based on the program parameters.

A “comparison window” as used herein is any number of consecutive positions that may compare a sequence to a reference sequence in the same number of consecutive positions after the two sequences have been optimally aligned. Includes a reference to one segment. In embodiments, the comparison window is about 20, 50, 100, 1000, 2500, 5000, 7500, 10000, 15000, 20000, 25000, or 30000, or at least about 20, 50, 100, 1000, 2500, 5000, 7500. 10000, 15000, 20000, 25000, or 30000 to about 31000, 32000, 33000, 34000 or 35000, about 31000, 32000, 33000, 34000 or 35000 or at least about 31000, 32000, 33000, 34000 or 35000 consecutive positions. including. In embodiments, the comparison window comprises about 20 or at least about 20 to about 31000, less than about 31000 or at least about 31000 consecutive positions. In embodiments, the comparison window comprises about 25,000 or at least about 25000 to about 31000, less than about 31000 or at least about 31000 consecutive positions. In embodiments, the comparison window comprises about 26000 or at least about 26000 to about 31000, less than about 31000 or at least about 31000 consecutive positions. In embodiments, the comparison window comprises about 27,000 or at least about 27000 to about 31000, less than about 31000 or at least about 31000 consecutive positions. In embodiments, the comparison window comprises about 28000 or at least about 28000 to about 31000, less than about 31000 or at least about 31000 consecutive positions. In embodiments, the comparison window comprises about 29000 or at least about 29000 to about 31000, less than about 31000 or at least about 31000 consecutive positions. In embodiments, the comparison window comprises about 30,000 or at least about 30,000 to about 31000, less than about 31000 or at least about 31000 consecutive positions. In embodiments, the comparison window comprises about 20 to about 600, about 50 to about 200, or about 100 to about 150 consecutive positions. In embodiments, the comparison window is the full length of the reference sequence, such as the sequence of the bacteriophage genome. Methods of aligning sequences for comparison are well known in the art. In an embodiment, the optimal alignment of sequences for comparison is described in, for example, Smith & Waterman, Adv. Appl. Math. 2:482 (1981) by the homology alignment algorithm of Needleman & Wunsch, J. Am. Mol. Biol. 48:443 (1970) by the homology alignment algorithm, Pearson & Lipman, Proc. Nat'l. Acad. Sci. USA 85:2444 (1988), by the similarity search method, these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, Wis., 575 Sc. Computerized alignment or by manual alignment and visual inspection (see, eg, Current Protocols in Molecular Biology (Ausubel et al., eds. 1995 addendum)).

Examples of suitable algorithms for determining percent sequence identity and sequence similarity are the BLAST and BLAST 2.0 algorithms, which are described in Altschul et al., Nuc. Acids Res. 25:3389-3402 (1977) and Altschul et al., J. Am. Mol. Biol. 215:403-410 (1990). As will be appreciated by those skilled in the art, software for performing BLAST analyzes is published through the website of the National Center for Biotechnology Information (NCBI). In embodiments, the parameters described herein are used to determine percent sequence identity for nucleic acids and proteins using BLAST and BLAST 2.0. In an embodiment, the BLAST algorithm comprises first identifying a high scoring sequence pair (HSP) by identifying a short word of length W in the query sequence, which word has the same length in the database sequence. When aligned with a word, it will match or meet some positive value threshold score T. In embodiments, T is referred to as the neighborhood word score threshold (Altschul et al., supra). In an embodiment, these initial neighborhood word hits act as seeds for initiating searches to find longer HSPs that contain them. In embodiments, word hits are extended in both directions along their respective sequences, as long as the cumulative alignment score can be increased. In an embodiment, the cumulative score uses the parameters M (reward score for a pair of matching residues; always >0) and N (penalty score for mismatching residues; always <0) for nucleotide sequences. Calculated. In embodiments, for amino acid sequences, a scoring matrix is used to calculate the cumulative score. In an embodiment, the extension of word hits in each direction reduces the cumulative alignment score by an amount X from its maximum achieved value; the cumulative score drops below zero due to the accumulation of one or more negative scoring residue alignments. Or it is stopped when the end of either sequence is reached. In an embodiment, the BLAST algorithm parameters W, T and X determine the sensitivity and speed of the alignment. In embodiments, the NCBI BLASTN or BLASTP programs are used to align sequences. In embodiments, the BLASTN or BLASTP programs use the defaults used by NCBI. In an embodiment, the BLASTN program (for nucleotide sequences) defaults: 28 word size (W); expected threshold (E) of 10; maximum match in query range set to 0; 1-2 match/ Use a mismatch score; a linear gap cost; a filter for the low complexity region used; and a mask only for the lookup table used. In embodiments, the BLASTP program (for amino acid sequences) defaults: word size (W) of 3; expected threshold (E) of 10; maximum match in query range set to 0; BLOSUM62 matrix (Henikoff & Henikoff, Proc. Natl. Acad. Sci. USA 89: 10915 (1992)); igistens gap cost: 11 and extension: 1; and conditional composition score matrix adjustment.

The terms “polypeptide”, “peptide” and “protein” are used interchangeably herein to refer to a polymer of amino acid residues. The term applies to amino acid polymers in which one or more amino acid residues is an artificial chemical mimetic of the corresponding naturally occurring amino acid, as well as naturally occurring and non-naturally occurring amino acid polymers.

The term "amino acid" refers to naturally occurring and synthetic amino acids, as well as amino acid analogs and amino acid mimetics that function in a manner similar to the naturally occurring amino acids. Naturally occurring amino acids are amino acids encoded by the genetic code, as well as amino acids that are subsequently modified, such as hydroxyproline, γ-carboxyglutamate and O-phosphoserine. Amino acid analogs are alpha carbons that have the same basic chemical structure as a naturally occurring amino acid, ie, hydrogen, carboxyl, amino and R groups such as homoserine, norleucine, methionine sulfoxide, methionine methylsulfonium. Is a compound having Such analogs have modified R groups (eg, norleucine) or modified peptide backbones, but retain the same basic chemical structure as a naturally occurring amino acid. Amino acid mimetics are chemical compounds that have a structure that is different from the general chemical structure of an amino acid, but that functions in a manner similar to a naturally occurring amino acid.

Amino acids may be referred to herein by either their commonly known three letter symbols or by the one-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission. Nucleotides may also be referred to by their commonly accepted one-letter code.

“Conservatively modified variants” applies to both amino acid and nucleic acid sequences. For a particular nucleic acid sequence, a conservatively modified variant is a nucleic acid that encodes the same or essentially the same amino acid sequence, or essentially the same sequence if the nucleic acid does not encode the amino acid sequence. Is. Due to the degeneracy of the genetic code, a large number of functionally identical nucleic acids encode any given protein. For example, the codons GCA, GCC, GCG and GCU all encode the amino acid alanine. Thus, at all positions where alanine is designated by a codon, the codon can be changed to any of the corresponding codons described without altering the encoded polypeptide. Such nucleic acid mutations are "silent mutations," which are a type of conservative modification mutation. Every nucleic acid sequence herein which encodes a polypeptide also describes every possible silent variation of the nucleic acid. Those skilled in the art will appreciate that each codon of a nucleic acid (except for AUG, which is usually the only codon for methionine, and TGG, which is usually the only codon for tryptophan), will result in a functionally identical molecule. Recognize that it is possible. Thus, each silent variation of a nucleic acid which encodes a polypeptide is implicit in each described sequence for expression products, but not for the actual probe sequence.

With respect to amino acid sequences, one of ordinary skill in the art would refer to individual substitutions of peptide, polypeptide, or protein sequences that change a single amino acid as "conservatively modified mutations" by which the amino acid is replaced with a chemically similar amino acid. We recognize that it is the body. Conservative substitution tables providing functionally similar amino acids are well known in the art. Such conservatively modified variants add to, but do not exclude, polymorphic variants, interspecies homologs and alleles.

The following eight groups each contain amino acids that are conservative substitutions for one another: 1) alanine (A), glycine (G); 2) aspartic acid (D), glutamic acid (E); 3) asparagine (N ), glutamine (Q); 4) arginine (R), lysine (K); 5) isoleucine (l), leucine (L), methionine (M), valine (V); 6) phenylalanine (F), tyrosine ( Y), tryptophan (W); 7) serine (S), threonine (T); and 8) cysteine (C), methionine (M) (see, for example, Creighton, Proteins (1984)).

The term “disease” refers to any deviation from the normal health of a mammal, including the condition in which disease symptoms are present, as well as deviations (eg, dysbiosis, infections, gene mutations, genetics). Defects, etc.), but symptoms are not yet apparent. In an embodiment, the disease is acne. In embodiments, the disease comprises cutaneous dysbiosis. In embodiments, the methods, compositions, systems, phages and probiotic bacteria provided herein include, but are not limited to, primates (such as humans), livestock, working animals and companion animals (eg, Suitable for use in subjects that are members of the class Vertebrate, Mammals, including companion animals. In embodiments, the subject is a human subject. As used herein, “symptoms” of a disease include, but are not limited to, the clinical or laboratory signs associated with the disease, which the subject can feel or observe.

As used herein, the term "skin dysbiosis" means a difference in skin microbiota as compared to a healthy or population. In embodiments, the dysbiosis is at the skin surface, within the skin (eg, within a skin area or layer of skin cells), within the gland, and/or within the pores of the skin. In embodiments, the dysbiosis is in sweat and/or sebum. In embodiments, the skin is on the face (eg, the subject's forehead, one or more cheeks, nose or chin). In embodiments, the skin is on the shoulders, chest or back. In embodiments, cutaneous dysbiosis comprises alterations in microbiome symbiont species diversity when compared to a healthy or population, reducing beneficial microbes and/or pathogenic symbionts (pathogenic or potentially Increased pathogenic microorganisms) and/or decreased overall flora species diversity. Many factors can cause dysbiosis, including hormonal changes (eg during adolescence), rare cleansing, cosmetic use, antibiotic use, psychological and physical stress, radiation and dietary changes ..

In embodiments, the composition is administered in a "therapeutically effective dose" to a subject suffering from acne. The effective amount for this use may depend on the severity of the disease and the general health of the patient. Single or multiple administrations of the compositions may be carried out depending on the dosage and the frequency required and tolerated by the patient. "Patient" or "subject" includes both human and other animals, especially mammals. Therefore, the method is applicable to both human therapy and veterinary applications.

"Pharmaceutically acceptable excipient" and "pharmaceutically acceptable carrier" means the administration of an active agent to and absorption by a subject without causing a significant adverse toxic effect on the patient. A substance that can be included in the composition of the present invention. Non-limiting examples of pharmaceutically acceptable excipients include water, NaCl, normal saline, Ringer's lactate, normal sucrose, normal glucose, binders, fillers, disintegrants, lubricants, coatings, sweeteners. Fragrances, fragrances, salt solutions (such as Ringer's solution), alcohols, oils, carbohydrates such as gelatin, lactose, amylose or starch, fatty acid esters, hydroxymethyl cellulose, polyvinylpyrrolidine and pigments and the like. . Such preparations should be sterilized and, where appropriate, lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts affecting osmotic pressure, buffers, coloring and/or aroma substances and the like. It is possible to mix such bacteriophages, probiotic bacteria and/or auxiliary compounds which do not deleteriously react with the compounds of the invention. One of ordinary skill in the art will recognize that other pharmaceutical excipients are useful in the present invention.

The term “contacting” may include reacting, interacting with, or physically contacting two chemical species with each other, eg, an enzyme described herein. And a biofilm containing an enzyme substrate. In another example, the two species can be cells of a bacteriophage and a bacterial species that the bacteriophage infects. In embodiments, contacting may include, for example, bacteriophage described herein. Including interacting with Acnes cells. In embodiments, contacting may include, for example, adding the enzymes described herein to P. Including interacting with the Acnes biofilm.

A "patient" or "subject in need of treatment" is a living member of the animal kingdom who has, or is at risk of having, the indicated disorder. In embodiments, the subject is a member of a species that includes individuals who naturally contract the disease. In embodiments, the subject is a mammal. Non-limiting examples of mammals include rodents (eg mouse and rat), primates (eg lemur, bush baby, monkey, ape and human), rabbits, dogs (eg companion dog, service dog or Includes police dogs, military dogs, work dogs such as race dogs or show dogs), cats (eg domestic cats), livestock (pigs, cows, donkeys, mules, bisons, goats, camels and sheep) and deer. In embodiments, the subject is a human.

The terms “subject”, “patient”, “individual”, etc. are not intended to be limiting and are generally interchangeable. That is, an "individual" described as a "patient" does not necessarily have a given disease, and may only seek a doctor's diagnosis.

As used herein, the abbreviation “sp.” for bacterial species refers to at least one bacterial species of the indicated genus (eg, 1, 2, 3, 4, 5 or more). There are many bacterial species). The abbreviation "spp." for bacterial species refers to two or more bacterial species of the indicated genus (eg, 2, 3, 4, 5, 6, 6, 7, 8, 9). One, ten or more). In embodiments, the methods and compositions provided herein are directed to a single genus within a designated genus or designated genera, or a designated genus or designated genus. Including two or more (eg, multiples containing more than two) species within the same genera. In embodiments, one, two, three, four, five or more or all of the indicated strains are isolated. In embodiments, the indicated bacterial species are administered together. In embodiments, each of the indicated strains is present in a single composition comprising each of the strains. In embodiments, each of the bacterial species is simultaneously, for example, about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 30 or 60, 1-5, 1-10, 1- 1 from each other. It is administered within 30, 1-60 or 5-15 seconds or minutes.

In this disclosure, "comprises," "comprising," "containing," "having," and the like are ascribed to these words in US Patent Law. It can have meaning, and can mean “includes,” “including,” and the like. Thus, the transitional term "comprising" is synonymous with "including," "containing," or "characterized by." , Or open-ended, and does not exclude additional, unlisted features, integers, steps, operations, elements and/or components. “Consisting essentially of” or “consisting essentially” also have the meaning ascribed to them in United States patent law, the term being open-ended and listed The presence of more than listed items permits the presence of more than listed items, but excludes prior art embodiments unless the basic or novel features of the item are modified by the presence of more items than listed. In contrast, the transitional phrase “consisting of” excludes any feature, integer, element, step, operation, component, and/or component not specified.

As used herein, the term “about” in the context of a numerical value or range is ±10% from the recited or claimed numerical value or range, unless the context requires a more limited range. Means

In the description and in the claims, terms such as "at least one of" or "one or more of" may precede the connection list of elements or features. The term "and/or" may also be present in a list of two or more elements or features. Unless the context in which the phrase is otherwise used is implicitly or clearly contradictory to another, such phrase may include any of the listed elements or features individually or on the listed elements or It is intended to mean any of the features in combination with any of the other listed elements or features. For example, the phrases "at least one of A and B", "one or more of A and B", and "A and/or B" each mean "A alone, B alone or A and B together". Is intended. A similar interpretation is intended for lists containing three or more items. For example, the phrases "at least one of A, B and C", "one or more of A, B and C" and "A, B and/or C" respectively refer to "A alone, B alone, C alone, A and B together, A and C together, B and C together, or A and B and C together”. Furthermore, the use of the term “based on” in this specification and claims is intended to mean “based, at least in part,” on the basis that features or elements not listed are also allowed. There is.

It is understood that where a parameter range is provided, all integers within that range and tenths thereof are also provided by the present invention. For example, "0.2 to 5 mg" is a disclosure including and including up to 5.0 mg of 0.2 mg, 0.3 mg, 0.4 mg, 0.5 mg, 0.6 mg, and the like.

As used in the description herein and throughout the claims that follow, the meanings of “a”, “an”, and “the” are other than context. Includes multiple references unless otherwise indicated by the method.

As used herein, "treating" or "treatment" of a condition, disease or disorder or of symptoms associated with a condition, disease or disorder is to obtain beneficial or desired results, including clinical results. It is an approach. A beneficial or desired clinical result is the reduction or amelioration of one or more symptoms or conditions, a diminished extent of a condition, disorder or disease, stabilization of a condition, disorder or disease state, onset of a condition, disorder or disease. Prevention, prevention of spread of a condition, disorder or disease, delay or slowing of progress of a condition, disorder or disease, delay or slowing of onset of a condition, disorder or disease, recovery or alleviation of a condition, disorder or disease state, and partial Whether or not, including but not limited to remission. “Treat” can also mean inhibiting the progression of a condition, disorder or disease, transiently slowing down the progression of a condition, disorder or disease, but in some cases the condition is , Permanently stopping the progression of the disorder or disease. When treating acne, the terms include, for example, cutaneous dysbiosis and/or cystic lesions, malignant tumors (closed and closed pores), blackheads (open and closed pores, exposure to air). Oils have a dark color, eg brown or black), mall red, tender bumps (pimples), pimples (pustules; papules with pimples on the tips), large, hard and painful lumps under the skin. It can be referred to as reducing the number or size of (nodules).

As used herein, the terms "treat" and "prevent" are not intended to be absolute terms. In embodiments, the treatment is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% of the severity of the disease, condition, or symptom of the disease or condition afflicted. Or it can refer to a 100% reduction. In embodiments, a method for treating a disease is considered to be treatment if there is a 10% reduction in one or more symptoms of the disease in the subject relative to the control. Thus, the reduction is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100% or 10% to 100% when compared to natural or control levels. Any percentage reduction of is possible. It is understood that treatment does not necessarily refer to the cure or complete resection of the disease, condition or symptoms of the disease or condition. In embodiments, reference to reducing, reducing or inhibiting is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% when compared to a control level. Including changes of 90% or greater, and such terms can include, but do not necessarily include, complete removal. Treatment can refer to any delay in onset, recovery of symptoms, improvement of the patient's skin appearance, and the like. The effect of treatment can be compared to an individual or pool of individuals who have not been treated or to the same patient at different times before or during treatment. In embodiments, the severity of the disease is reduced, for example, by at least 10% when compared to pre-administered individuals or untreated control individuals. In some aspects, the severity of the disease is reduced by at least 25%, 50%, 75%, 80% or 90%, or in some cases no longer detectable using standard diagnostic techniques Is. In embodiments, the treatment is effective in reducing at least one symptom of acne. In embodiments, the treatment is effective to reduce the level of pimples (pustules) on the face, forehead, chest, back and/or shoulders of the subject. In embodiments, the treatment is effective to reduce the level of malignant tumor (closed and closed pores) on the face, forehead, chest, back and/or shoulders of the subject. In embodiments, the treatment is effective to reduce the level of blackheads (open clogged pores) on the subject's face, forehead, chest, back and/or shoulders. In embodiments, the treatment is effective to reduce the level of papules on the subject's face, forehead, chest, back and/or shoulders. In embodiments, the treatment is effective to reduce the level of subsurface hard and painful lumps (nodules) of the skin of the subject's face, forehead, chest, back and/or shoulders. In embodiments, the treatment is effective to reduce the level of cystic lesions on the face, forehead, chest, back and/or shoulders of the subject. In embodiments, the level (eg, number) is reduced as compared to before treatment began. In embodiments, the level (eg, number) is reduced compared to a corresponding subject who has suffered from acne but has not received treatment. In embodiments, the level (eg, number) is reduced as compared to a corresponding subject who has suffered from acne but has not received treatment containing bacteriophage.

The terms "effective amount", "effective dose", "therapeutically effective amount" and the like refer to an amount of an agent sufficient to ameliorate the disorders described herein. For example, for a given parameter, a therapeutically effective amount is at least 5%, 10%, 15%, 20%, 25%, 40%, 50%, 60%, 75%, 80%, 90% or at least 100%. Indicates an increase or decrease in The therapeutic effect can also be expressed as a "fold" increase or decrease. For example, a therapeutically effective amount can have an effect of at least 1.2-fold, 1.5-fold, 2-fold, 5-fold or more over the control.

The term "diagnosis" refers to the relative probability that a subject has a given metabolic disorder. Symptoms and diagnostic criteria are summarized herein. Similarly, the term "prognosis" refers to the relative probability that a particular future outcome will occur in a subject. For example, in the context of the present invention, prognosis can refer to the likelihood that an individual will develop acne.
Prognosis can refer to the likely severity of the disease (eg, severity of symptoms, rate of functional decline, etc.). As one of ordinary skill in the medical diagnostic arts will recognize, the term is not intended to be absolute.

Compositions and Combination Embodiments Comprising Bacteriophage In at least one P. Provided herein is a composition comprising, consisting essentially of, or consisting of an acnes bacteriophage, at least one anti-acne compound and a pharmaceutically acceptable carrier.

In an aspect, provided herein is a composition comprising at least one Propionibacterium acnes bacteriophage, no more than one anti-acne compound and a pharmaceutically acceptable carrier.

In embodiments, provided herein is a composition comprising an active ingredient consisting of at least one Propionibacterium acnes bacteriophage and one or less anti-acne compound and a pharmaceutically acceptable carrier.

In an embodiment, at least one P. Provided herein are compositions comprising acnes bacteriophage, at least one anti-acne compound and a pharmaceutically acceptable carrier, and free of probiotic bacteria.

In embodiments, the at least one anti-acne compound is benzoyl peroxide. In embodiments, benzoyl peroxide is present at a concentration of 2.5%-10% (weight/volume). In embodiments, benzoyl peroxide is present at a concentration of less than 2.5% but greater than about 0.1%, 0.5%, 1%, 1.5% or 2% (weight/volume). ing. In embodiments, benzoyl peroxide is 2.5% to 10%, such as about 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%. %, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5% or 10% (weight/volume). In embodiments, benzoyl peroxide is present at a concentration of less than 2.5% but greater than about 0.1%, 0.5%, 1%, 1.5% or 2% (weight/volume). ing.

In embodiments, the at least one anti-acne compound is salicylic acid. In embodiments, salicylic acid is present at a concentration of 0.5%-2% (weight/volume). In embodiments, salicylic acid is present at a concentration of less than 0.5% but greater than about 0.1% (weight/volume). In embodiments, salicylic acid is 0.5% to 2%, such as about 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.1%. , 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9% or 2% (weight/volume) concentration Exists in. In embodiments, salicylic acid is present at a concentration of less than 0.5% but greater than about 0.1% (weight/volume).

In embodiments, the at least one anti-acne compound is sulfur. In embodiments, sulfur is present at a concentration of 3%-10% (weight/volume). In embodiments, sulfur is less than 3% but is present at a concentration greater than about 0.1%, 0.5%, 1%, 1.5%, 2% or 2.5% (weight/volume). doing. In embodiments, sulfur is 3% to 10%, such as about 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%. , 7.5%, 8%, 8.5%, 9%, 9.5% or 10% (weight/volume). In embodiments, sulfur is less than 3% but is present at a concentration greater than about 0.1%, 0.5%, 1%, 1.5%, 2% or 2.5% (weight/volume). doing. In embodiments, resorcinol is present at a concentration of 2% and sulfur is between 3% and 8% (eg, about 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5%, 0.5%, 6%, 6.5%, 7%, 7.5% or 8%) (weight/volume).

In embodiments, the at least one anti-acne compound is resorcinol and sulfur. In an embodiment, resorcinol is present at a concentration of 2% and sulfur is present at a concentration of 3%-8% (weight/volume). In embodiments, resorcinol is present at a concentration of 2% and sulfur is between 3% and 8% (eg, about 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5%, 0.5%, 6%, 6.5%, 7%, 7.5% or 8%) (weight/volume).

In embodiments, the at least one anti-acne compound comprises resorcinol monoacetate and sulfur. In an embodiment, resorcinol monoacetate is present at a concentration of 3% and sulfur is present at a concentration of 3% to 8% (weight/volume). In an embodiment, resorcinol monoacetate is present at a concentration of 3% and sulfur is between 3% and 8% (eg, about 2.5%, 3%, 3.5%, 4%, 4.5%, 5%. 5.5%, 6%, 6.5%, 7%, 7.5% or 8%) (weight/volume).

In an embodiment, P. Acnes bacteriophage is about 1×10 ⁶ , 2×10 ⁶ , 3×10 ⁶ , 4×10 ⁶ , 5×10 ⁶ , 6×10 ⁶ , 7×10 ⁶ , 8×10 ⁶ , 9×10 ^6. 1×10 ⁷ , 2×10 ⁷ , 3×10 ⁷ , 4×10 ⁷ , 5×10 ⁷ 6×10 ⁷ , 7×10 ⁷ , 8×10 ⁷ , 9×10 ⁷ , 1×10 ⁸ , 2x10 ⁸ , 3x10 ⁸ , 4x10 ⁸ , 5x10 ⁸ , 6x10 ⁸ , 7x10 ⁸ , 8x10 ⁸ , 9x10 ⁸ , 1x10 ⁹ , 2x10 ⁹ , 3×10 ⁹ , 4×10 ⁹ , 5×10 ⁹ , 6×10 ⁹ , 7×10 ⁹ , 8×10 ⁹ , 9×10 ⁹ , 1×10 ¹⁰ , 2×10 ¹⁰ , 3×10 ¹⁰ , It is present in an amount of 4×10 ¹⁰ , 5×10 ¹⁰ , 6×10 ¹⁰ , 7×10 ¹⁰ , 8×10 ¹⁰ , 9×10 ¹⁰ , or 1×10 ¹¹ plaque forming units (pfu). In an embodiment, P. Acnes bacteriophage is present in an amount of about 1×10 ⁶ to 1×10 ¹¹ pfu. In an embodiment, P. Acnes bacteriophage may be about 1×10 ⁶ to 1×10 ⁸ , about 1×10 ⁸ to 1×10 ⁹ , about 1×10 ⁹ to 1×10 ¹⁰ , about 1×10 ⁹ to 1×10 ¹¹ or about. It is present in an amount of 1×10 ¹⁰ to 1×10 ¹¹ pfu.

In embodiments, the probiotic bacterium is about 1×10 ⁶ , 2×10 ⁶ , 3×10 ⁶ , 4×10 ⁶ , 5×10 ⁶ , 6×10 ⁶ , 7×10 ⁶ , 8×10 ^6. , 9×10 ⁶ , 1×10 ⁷ , 2×10 ⁷ , 3×10 ⁷ , 4×10 ⁷ , 5×10 ⁷ 6×10 ⁷ , 7×10 ⁷ , 8×10 ⁷ , 9×10 ⁷ , 1×10 ⁸ , 2×10 ⁸ , 3×10 ⁸ , 4×10 ⁸ , 5×10 ⁸ , 6×10 ⁸ , 7×10 ⁸ , 8×10 ⁸ , 9×10 ⁸ , 1×10 ⁹ , 2x10 ⁹ , 3x10 ⁹ , 4x10 ⁹ , 5x10 ⁹ , 6x10 ⁹ , 7x10 ⁹ , 8x10 ⁹ , 9x10 ⁹ , 1x10 ¹⁰ , 2x10 ¹⁰ , Present in an amount of 3×10 ¹⁰ , 4×10 ¹⁰ , 5×10 ¹⁰ , 6×10 ¹⁰ , 7×10 ¹⁰ , 8×10 ¹⁰ , 9×10 ¹⁰ , or 1×10 ¹¹ colony forming units (cfu). doing. In an embodiment, the probiotic bacterium is present in an amount of about 1×10 ⁶ to 1×10 ¹¹ cfu. In embodiments, the probiotic bacterium is about 1×10 ⁶ to 1×10 ⁸ , about 1×10 ⁸ to 1×10 ⁹ , about 1×10 ⁹ to 1×10 ¹⁰ , about 1×10 ⁹ to 1. is present in an amount of × ^{10 11} or about ^{1 × 10 10 ~1 × 10 11} cfu.

In embodiments, the anti-acne compound is an antibiotic, retinoid or alpha hydroxy acid.

In an embodiment, P. Provided herein is a composition comprising an Acnes bacteriophage and an enzyme.

In an embodiment, at least one P. Acnes bacteriophage, a combination comprising, consisting essentially of, or consisting of at least one anti-acne compound, wherein the at least one P. Provided herein are combinations of Acnes bacteriophage and at least one anti-acne compound in a composition that further comprises a pharmaceutically acceptable carrier.

In an embodiment, P. Provided herein are combinations that include an Acnes bacteriophage and an enzyme.

In an embodiment, P. Acnes bacteriophage has a linear double-stranded DNA genome.

In an embodiment, P. Acnes bacteriophages are within the family of bacteriophage Siphoviridae.

In an embodiment, the bacteriophage is a wild type bacteriophage. In embodiments, the bacteriophage comprises at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90% of the genomic sequence of the wild-type Acnes bacteriophage. %, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99 Have a genome with nucleotide sequences that are 0.5%, 99.6%, 99.7%, 99.8% or 99.9% identical. Non-limiting examples of wild-type Acnes bacteriophage genomic sequences are as follows.

In embodiments, the bacteriophage is the bacteriophage deposited under accession number NCIMB 41349, 41350 or 41351. In embodiments, the bacteriophage is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88 in the genome of the bacteriophage deposited under accession number NCIMB 41349. %, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.1%, 99.2%, 99.3%, It has a genome with nucleotide sequences that are 99.4%, 99.5%, 99.6%, 99.7%, 99.8% or 99.9% identical. In an embodiment, the bacteriophage is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88 in the genome of the bacteriophage deposited under accession number NCIMB 41350. %, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.1%, 99.2%, 99.3%, It has a genome with nucleotide sequences that are 99.4%, 99.5%, 99.6%, 99.7%, 99.8% or 99.9% identical. In embodiments, the bacteriophage is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88 in the genome of the bacteriophage deposited under accession number NCIMB 41351. %, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.1%, 99.2%, 99.3%, It has a genome with nucleotide sequences that are 99.4%, 99.5%, 99.6%, 99.7%, 99.8% or 99.9% identical.

In an embodiment, the bacteriophage has at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90% of the nucleotide sequence of SEQ ID NO:1. 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5. Have a genome with nucleotide sequences that are %, 99.6%, 99.7%, 99.8% or 99.9% identical. In an embodiment, the bacteriophage has a genome with a nucleotide sequence that is identical to the nucleotide sequence of SEQ ID NO:1.

In an embodiment, the bacteriophage genome is 5′ to 3′ end, small terminase, large terminase, portal protein, gp4, scaffold protein, major head protein, gp7, gp8, gp9, gp10, major tail protein, gp12, gp13, tape measure protein, small tail subunit, optionally protease, gp17, gp18, tail protein, amidase, holin, gp22, gp23, sigma factor, gp25, gp26, gp27, gp28, gp29, gp30, DNA It encodes a primase, a DNA primase 2, gp33, a DNA helicase, gp35, gp36, an exonuclease, p38, gp39, gp40, gp41, gp42, gp43, gp44, gp45, gp46, gp47 and gp48.

In an embodiment, the composition is P. It further comprises an Acnes biofilm degrading enzyme.

In embodiments, the enzyme is an anti-aging enzyme. In embodiments, the anti-aging enzyme is superoxide dismutase or peroxidase.

In an embodiment, the enzyme is P. Acnes is a biofilm degrading enzyme. In embodiments, the enzyme is a glycosidase, protease, deoxyribonuclease or restriction endonuclease. In embodiments, the enzyme is glycosidase. In embodiments, the glycosidase is a glycoside hydrolase. In embodiments, the enzyme catalyzes the hydrolysis of a linear polymer of N-acetyl-D-glucosamine. In an embodiment, the enzyme is β-hexosaminidase. In embodiments, the enzyme hydrolyzes the β-1,6-glycosidic bond of the acetylglucosamine polymer. In embodiments, the enzyme is deoxyribonuclease I, a restriction endonuclease, papain, bromelain, trypsin, proteinase K, subtilisin, seratiopeptidase, dispascin, alginate lyase, amylase or cellulase. In an embodiment, the enzyme is Dispersin B. In embodiments, the enzyme is a protease and the protease is proteinase K or subtilisin.

In an embodiment, the enzyme is dispersin. In an embodiment, the enzyme is Dispersin B. In embodiments, the enzyme retains enzymatic activity (eg, at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity as compared to the native protein. In) naturally occurring forms, homologues, isoforms or variants of Dispersin (such as Dispersin B). In embodiments, the variant is at least 90%, 95%, 96% over the entire sequence or a portion of the sequence (eg, 50, 100, 150 or 200 contiguous amino acid moieties) as compared to the naturally occurring form. It has 97%, 98%, 99% or 100% amino acid sequence identity. A non-limiting example of a DNA sequence encoding Dispersin B is as follows.

Non-limiting examples of Dispersin B amino acid sequences are as follows.

In an embodiment, the enzyme is alginate lyase. In embodiments, the enzyme maintains the enzymatic activity of alginate lyase (eg, at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% compared to the native protein. % Of activity) naturally occurring forms, homologues, isoforms or variants of alginate lyase. In embodiments, the variant is at least 90%, 95%, 96% over the entire sequence or a portion of the sequence (eg, 50, 100, 150 or 200 contiguous amino acid moieties) as compared to the naturally occurring form. It has 97%, 98%, 99% or 100% amino acid sequence identity. Non-limiting examples of DNA sequences encoding alginate lyase are as follows.

Non-limiting examples of alginate lyase amino acid sequences are as follows.

In embodiments, the enzyme is amylase. In embodiments, the enzyme maintains the enzymatic activity of amylase (eg, at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% relative to the native protein). Activity) a naturally occurring form, homologue, isoform or variant of amylase. In embodiments, the variant is at least 90%, 95%, 96% over the entire sequence or a portion of the sequence (eg, 50, 100, 150 or 200 contiguous amino acid moieties) as compared to the naturally occurring form. It has 97%, 98%, 99% or 100% amino acid sequence identity. Non-limiting examples of DNA sequences encoding amylase are as follows.

Non-limiting examples of amylase amino acid sequences are as follows.

In embodiments, the enzyme is cellulase. In embodiments, the enzyme maintains the enzymatic activity of cellulase (eg, at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% relative to the native protein). (Within the activity of) cellulase is a naturally occurring form, homologue, isoform or variant. In embodiments, the variant is at least 90%, 95%, 96% over the entire sequence or a portion of the sequence (eg, 50, 100, 150 or 200 contiguous amino acid moieties) as compared to the naturally occurring form. It has 97%, 98%, 99% or 100% amino acid sequence identity. Non-limiting examples of DNA sequences encoding cellulase are as follows.

Non-limiting examples of cellulase amino acid sequences are as follows.

In an embodiment, the enzyme is proteinase K. In embodiments, the enzyme maintains the enzymatic activity of proteinase K (eg, at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100 compared to the native protein. % Of activity) is a naturally occurring form, homologue, isoform or variant of proteinase K. In embodiments, the variant is at least 90%, 95%, 96% over the entire sequence or a portion of the sequence (eg, 50, 100, 150 or 200 contiguous amino acid moieties) as compared to the naturally occurring form. It has 97%, 98%, 99% or 100% amino acid sequence identity. A non-limiting example of a DNA sequence encoding proteinase K is as follows.

Non-limiting examples of proteinase K amino acid sequences are as follows.

In an embodiment, the enzyme is subtilisin. In embodiments, the enzyme maintains the enzymatic activity of subtilisin (eg, at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% relative to the native protein. (Within the activity of) subtilisin is a naturally occurring form, homologue, isoform or variant. In embodiments, the variant is at least 90%, 95%, 96% over the entire sequence or a portion of the sequence (eg, 50, 100, 150 or 200 contiguous amino acid moieties) as compared to the naturally occurring form. It has 97%, 98%, 99% or 100% amino acid sequence identity. A non-limiting example of a DNA sequence encoding subtilisin is as follows.

Non-limiting examples of subtilisin amino acid sequences are as follows.

In an embodiment, the enzyme is trypsin. In embodiments, the enzyme maintains the enzymatic activity of trypsin (eg, at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% relative to the native protein). (Within the activity of) trypsin is a naturally occurring form, homologue, isoform or variant. In embodiments, the variant is at least 90%, 95%, 96% over the entire sequence or a portion of the sequence (eg, 50, 100, 150 or 200 contiguous amino acid moieties) as compared to the naturally occurring form. It has 97%, 98%, 99% or 100% amino acid sequence identity. Non-limiting examples of trypsin-encoding DNA sequences are as follows.

Non-limiting examples of trypsin amino acid sequences are as follows.

In an embodiment, the enzyme is seratiopeptidase. In embodiments, the enzyme maintains the enzymatic activity of serathiopeptidase (eg, at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or as compared to the native protein). (Within 100% activity) is a naturally occurring form, homologue, isoform or variant of serathiopeptidase. In embodiments, the variant is at least 90%, 95%, 96% over the entire sequence or a portion of the sequence (eg, 50, 100, 150 or 200 contiguous amino acid moieties) as compared to the naturally occurring form. It has 97%, 98%, 99% or 100% amino acid sequence identity. A non-limiting example of a DNA sequence encoding a seratiopeptidase is as follows.

A non-limiting example of a seratiopeptidase amino acid sequence is as follows.

In embodiments, the enzyme is deoxyribonuclease. In embodiments, the enzyme maintains the enzymatic activity of a deoxyribonuclease (eg, at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100 compared to the native protein. % Of activity) naturally occurring forms, homologues, isoforms or variants of deoxyribonuclease. In embodiments, the variant is at least 90%, 95%, 96% over the entire sequence or a portion of the sequence (eg, 50, 100, 150 or 200 contiguous amino acid moieties) as compared to the naturally occurring form. It has 97%, 98%, 99% or 100% amino acid sequence identity. In an embodiment, the enzyme is Deoxyribonuclease I. In an embodiment, the deoxyribonuclease I is bovine pancreatic desoxyribonuclease I. A non-limiting example of a DNA sequence encoding bovine pancreatic desoxyribonuclease I is as follows.

A non-limiting example of a bovine pancreatic deoxyribonuclease I amino acid sequence is as follows.

In embodiments, the composition or combination comprises probiotic bacteria.

In an embodiment, the probiotic bacterium is P. Genus species (P. sp.), Staphylococcus sp. and/or Corynebacterium sp. bacteria.

In an embodiment, the probiotic bacterium is a bacterium within the Betaproteobacteria class.

In an embodiment, the probiotic bacterium is a probiotic P. It is Acnes.

In an embodiment, P. Acnes has (a) a 16S ribosomal DNA (rDNA) sequence with a T992C mutation as compared to the KPA 171202 reference strain 16S rDNA sequence shown as SEQ ID NO: 2; (b) KPA 171202 reference strain 16S shown as SEQ ID NO: 2 having a 16S rDNA sequence with the T838C mutation as compared to the rDNA sequence; (c) having a 16S rDNA sequence with the C1322T mutation as compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2; (d) Has a 16S rDNA sequence with a C986T mutation as compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO: 2; (e) has a 16S rDNA sequence identical to the sequence of SEQ ID NO: 3; (f) sequence Having a 16S rDNA sequence identical to the sequence of number 4; (g) without a linear plasmid; (h) without a plasmid containing a virulence factor; and/or (i) an extrachromosomal lipase and/or There is no plasmid encoding the adherent virulence factor.

In an embodiment, P. Acnes bacterium (a) is a pathogenic P. cerevisiae when grown in suspension culture. About 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60% of the level of lipase produced by the Acnes strain Produces less than 65%, 70%, 75%, 80%, 85%, 90% or 95%; (b) Pathogenic P. when grown in adherent culture. About 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60% of the level of lipase produced by the Acnes strain Produces less than 65%, 70%, 75%, 80%, 85%, 90% or 95%; (c) Adhesion to epithelial cells causes pathogenic P. At least 50% less than the Acnes strain; and/or (d) pathogenic P. Less inflammatory than Acnes strain.

In embodiments, the combination or composition comprises at least one additional probiotic bacterium. In an embodiment, the at least one additional probiotic bacterium comprises Propionibacterium granulosum and/or Propionibacterium avidum.

In an embodiment, the pathogenic P. The Acnes strain has (a) a 16S rDNA sequence with the G1058C mutation as compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2; (b) Compared to the KPA1712022 reference strain 16S rDNA sequence shown as SEQ ID NO:2 Has a 16S rDNA sequence with G1058C and A1201C; (c) has a 16S rDNA sequence with a G529A mutation as compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2; (d) as SEQ ID NO:2 16S rDNA sequence with G1004A and T1007C mutations compared to the KPA171202 reference strain 16S rDNA sequence shown; (e) 16S rDNA with G1268A mutation compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2 Having a sequence; (f) having a 16S rDNA sequence with the T554C and G1058C mutations compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2; (g) a 16S identical to the sequence of SEQ ID NO:5 having a rDNA sequence; (h) having a 16S rDNA sequence identical to the sequence of SEQ ID NO:6; (i) having a 16S rDNA sequence identical to the sequence of SEQ ID NO:7; (j) a SEQ ID NO:8 A 16S rDNA sequence that is identical to the sequence of SEQ ID NO:; (k) has a 16S rDNA sequence that is identical to the sequence of SEQ ID NO:9; and/or (l) is a 16S rDNA sequence that is identical to the sequence of SEQ ID NO:10 Have.

In embodiments, the combination or composition comprises at least one additional P. Further includes acnes bacteriophage.

In embodiments, the combination or composition further comprises a pharmaceutically acceptable carrier. In embodiments, the pharmaceutically acceptable carrier comprises an emulsion. In embodiments, the emulsion is an oil-in-water emulsion or a water-in-oil emulsion. In embodiments, the combination or composition comprises or is in the form of a cream, lotion, suspension or aqueous solution.

In embodiments, compositions comprising bacteriophage are provided. In embodiments, the composition is formulated for topical application to the skin (ie, the composition is a topical composition). In embodiments, the composition is a pharmaceutical composition.

In an embodiment, wild type P. Acnes bacteriophage and isolated probiotic P. There is provided a pharmaceutical composition comprising Acnes bacteria. In embodiments, the composition further comprises a pharmaceutically acceptable carrier.

In an embodiment, bacteriophage and/or isolated probiotic P. There is provided a pharmaceutical composition comprising A. acnes and a pharmaceutically acceptable carrier.

In embodiments, the pharmaceutical composition is formulated for topical administration to the skin. In embodiments, the pharmaceutically acceptable carrier comprises an emulsion. In embodiments, the emulsion is an oil-in-water emulsion or a water-in-oil emulsion. In embodiments, the pharmaceutical composition is in the form of a cream, lotion, suspension or aqueous solution.

In embodiments, the composition or combination has at least about 2, 3, 4, 5, 6, 7, 8, 9 or 10 P. Includes acnes bacteriophage. In an embodiment, P. Acnes bacteriophages are more than one type of P. Includes acnes bacteriophage.

In an embodiment, the isolated probiotic P. The combination or composition comprising the bacterium Acnes may further comprise at least one additional bacterium.

In an embodiment, P. Acnes has a 16S rDNA sequence containing the T992C, T838C, C1322T and/or C986T mutations compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2. In an embodiment, P. Acnes contains a 16S rDNA sequence with the T838C and C1322T mutations as compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2. In an embodiment, P. Acnes is ProI strain. In an embodiment, P. Acnes contains a 16S rDNA sequence with the C986T and T992C mutations as compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2. In an embodiment, P. Acnes is a ProII strain. In an embodiment, P. Acnes contains (a) a 16S rDNA sequence with the T992C mutation as compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO: 2; (b) a KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO: 2 In comparison, it contains a 16S rDNA sequence with the T838C mutation; (c) contains a 16S rDNA sequence with the C1322T mutation compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2; (d) SEQ ID NO:2. A 16S rDNA sequence with a C986T mutation as compared to the KPA171202 reference strain 16S rDNA sequence shown as; (e) containing a 16S rDNA sequence identical to the sequence of SEQ ID NO:3; (f) a sequence of SEQ ID NO:4 A 16S rDNA sequence that is identical to;; (g) no linear plasmid; (h) no plasmid containing virulence factors; and/or (i) extrachromosomal lipase and/or adherent virulence factors It does not include a plasmid encoding In an embodiment, P. Acnes has a 16S rDNA which is identical to the sequence SEQ ID NO: 3 or 4.

In an embodiment, P. Acnes bacterium (a) is a pathogenic P. cerevisiae when grown in suspension culture. About 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60% of the level of lipase produced by the Acnes strain Produces less than 65%, 70%, 75%, 80%, 85%, 90% or 95%; (b) Pathogenic P. when grown in adherent culture. About 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60% of the level of lipase produced by the Acnes strain Produces less than 65%, 70%, 75%, 80%, 85%, 90% or 95%; (c) Adhesion to epithelial cells causes pathogenic P. At least 50% less than the Acnes strain; and/or (d) pathogenic P. Less inflammatory than Acnes strain. In an embodiment, the pathogenic P. The Acnes strain has (a) a 16S rDNA sequence with the G1058C mutation as compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2; (b) Compared to the KPA1712022 reference strain 16S rDNA sequence shown as SEQ ID NO:2 Has a 16S rDNA sequence with G1058C and A1201C; (c) has a 16S rDNA sequence with a G529A mutation as compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2; (d) as SEQ ID NO:2 16S rDNA sequence with G1004A and T1007C mutations compared to the KPA171202 reference strain 16S rDNA sequence shown; (e) 16S rDNA with G1268A mutation compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2 Having a sequence; (f) having a 16S rDNA sequence with the T554C and G1058C mutations compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2; (g) a 16S identical to the sequence of SEQ ID NO:5 having a rDNA sequence; (h) having a 16S rDNA sequence identical to the sequence of SEQ ID NO:6; (i) having a 16S rDNA sequence identical to the sequence of SEQ ID NO:7; (j) a SEQ ID NO:8 A 16S rDNA sequence that is identical to the sequence of SEQ ID NO:; (k) has a 16S rDNA sequence that is identical to the sequence of SEQ ID NO:9; and/or (l) is a 16S rDNA sequence that is identical to the sequence of SEQ ID NO:10 Have.

SEQ ID NO:2 is a 16S rDNA sequence representing the KPA171202 reference strain and is as follows:

SEQ ID NO:3 is the 16S rDNA sequence representing the ProI probiotic strain and is as follows:
Nucleotide 838. ． 838
ProI mutation T838C
Nucleotide 1322. ． 1322
ProI mutation C1322T

SEQ ID NO:4 is the 16S rDNA sequence representing the ProII probiotic strain and is as follows:
Nucleotide 986. ． 986
ProII mutation C986T
Nucleotide 992. ． 992
ProII mutation T992C

In an embodiment, P. Acnes is a virulent P. aeruginosa when grown in suspension culture. About 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60% of the level of lipase produced by the Acnes strain It produces less than 65%, 70%, 75%, 80%, 85%, 90% or 95%. In an embodiment, P. Acnes is a virulent P. aeruginosa when grown in suspension culture. About 1-5%, 1-10%, 1-20%, 1-30%, 5-50%, 5-40%, 5-30%, 5-20% of the level of lipase produced by the Acnes strain 5-10%, 10-50%, 10-40%, 10-30%, 10-20%, 20-50%, 20-40%, or 20-30%. In an embodiment, P. Acnes is a virulent P. aeruginosa when grown in suspension culture. It produces less than about 5% of the level of lipase produced by the Acnes strain. In an embodiment, P. Acnes is a virulent P. aeruginosa when grown in suspension culture. It produces less than about 10% of the level of lipase produced by the Acnes strain. In an embodiment, P. Acnes is a virulent P. aeruginosa when grown in suspension culture. It produces less than about 20% of the level of lipase produced by the Acnes strain. In an embodiment, P. Acnes is a virulent P. aeruginosa when grown in suspension culture. It produces less than about 30% of the level of lipase produced by the Acnes strain. In an embodiment, P. Acnes is a virulent P. aeruginosa when grown in suspension culture. It produces less than about 40% of the level of lipase produced by the Acnes strain. In an embodiment, P. Acnes is a virulent P. aeruginosa when grown in suspension culture. It produces less than about 50% of the level of lipase produced by the Acnes strain. In an embodiment, P. Acnes is a pathogenic P. cerevisiae when grown in adherent culture. About 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60% of the level of lipase produced by the Acnes strain It produces less than 65%, 70%, 75%, 80%, 85%, 90% or 95%. In an embodiment, P. Acnes is a pathogenic P. cerevisiae when grown in adherent culture. It produces less than about 5% of the level of lipase produced by the Acnes strain. In an embodiment, P. Acnes is a pathogenic P. cerevisiae when grown in adherent culture. It produces less than about 10% of the level of lipase produced by the Acnes strain. In an embodiment, P. Acnes is a pathogenic P. cerevisiae when grown in adherent culture. It produces less than about 20% of the level of lipase produced by the Acnes strain. In an embodiment, P. Acnes is a pathogenic P. cerevisiae when grown in adherent culture. It produces less than about 30% of the level of lipase produced by the Acnes strain. In an embodiment, P. Acnes is a pathogenic P. cerevisiae when grown in adherent culture. It produces less than about 40% of the level of lipase produced by the Acnes strain. In an embodiment, P. Acnes is a pathogenic P. cerevisiae when grown in adherent culture. It produces less than about 50% of the level of lipase produced by the Acnes strain. In an embodiment, P. Acnes is a pathogenic P. cerevisiae when grown in adherent culture. About 1-5%, 1-10%, 1-20%, 1-30%, 5-50%, 5-40%, 5-30%, 5-20% of the level of lipase produced by the Acnes strain 5-10%, 10-50%, 10-40%, 10-30%, 10-20%, 20-50%, 20-40%, or 20-30%. In embodiments, the lipase is an extracellular lipase.

In an embodiment, P. The level of lipase produced by a Bacillus acnes (eg, probiotic or pathogenic P. acnes, for comparison) is the level of lipase in the culture supernatant. In embodiments, the culture supernatant is filtered. In embodiments, the culture supernatant is from a liquid (floating) culture. In embodiments, the culture supernatant is from adherent culture. Non-limiting examples of methods for detecting levels of lipase include absorbance, Bradford protein assay, Burette test derived assay, fluoresamine, amino black, colloidal gold, nitrogen detection, high performance liquid chromatography (HPLC), liquid chromatography. -Includes mass spectrometry (LC/MS), enzyme linked immunosorbent assay (ELISA), protein immunoprecipitation, immunoelectrophoresis and Western blot.

In an embodiment, P. The bacterium Acnes causes pathogenic P. At least about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70% more than Acnes strains , 75%, 80%, 85%, 90% or 95% less. In an embodiment, P. The bacterium Acnes causes pathogenic P. At least about 50% less than the Acnes strain. In an embodiment, P. The bacterium Acnes causes pathogenic P. At least about 60% less than the Acnes strain. In an embodiment, P. The bacterium Acnes causes pathogenic P. At least about 70% less than the Acnes strain. In an embodiment, P. The bacterium Acnes causes pathogenic P. At least about 80% less than the Acnes strain. In an embodiment, P. The bacterium Acnes causes pathogenic P. At least about 90% less than the Acnes strain. In an embodiment, P. The bacterium Acnes causes pathogenic P. 1-5%, 1-10%, 1-20%, 1-30%, 5-50%, 5-40%, 5-30%, 5-20%, 5-10%, more than Acnes strain -50%, 10-40%, 10-30%, 10-20%, 20-50%, 20-40%, or 20-30%, 50-60, 50-70, 50-80, 50-90. , 60-80, 70-90 less.

In an embodiment, P. Adhesion of A. acnes (eg, probiotic or pathogenic P. acnes, as for comparison) is determined using A-432 epithelial cells. In embodiments, epithelial cells are confluent in tissue culture plates or flasks. In an embodiment, the adhesion is P. cerevisiae adherent to cultured epithelial cells. Detected by determining the number of colonies formed by the bacterium Acnes.

In an embodiment, P. Acnes bacteria are pathogenic P. Less inflammatory than Acnes strain.

In an embodiment, the pathogenic P. Acnes strain or pathogenic P. Lower levels of inflammatory cytokines (eg, at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or less than the compounds produced by the bacteria of the Acnes strain). 90% less) Acnes or P. If released by immune cells in contact with the compound produced by P. acnes, P. Acnes bacteria are pathogenic P. Less inflammatory than Acnes strain. In embodiments, lower levels of inflammatory cytokines are associated with P. If it is released in tissues that are in contact with B. acnes (such as skin tissue), P. Acnes bacteria are pathogenic P. Less inflammatory than Acnes strain. In embodiments, the tissue is skin tissue. In an embodiment, the tissue is, for example, mouse ear tissue. In embodiments, the inflammatory cytokine is IL-1β, IL-6, IL-17 or TNFα, or any combination thereof.

In an embodiment, the pathogenic P. The Acnes strain has (a) a 16S rDNA sequence with the G1058C mutation as compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2; (b) Compared to the KPA1712022 reference strain 16S rDNA sequence shown as SEQ ID NO:2 Has a 16S rDNA sequence with G1058C and A1201C; (c) has a 16S rDNA sequence with a G529A mutation as compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2; (d) as SEQ ID NO:2 16S rDNA sequence with G1004A and T1007C mutations compared to the KPA171202 reference strain 16S rDNA sequence shown; (e) 16S rDNA with G1268A mutation compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2 Having a sequence; (f) having a 16S rDNA sequence with the T554C and G1058C mutations compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2; (g) a 16S identical to the sequence of SEQ ID NO:5 having a rDNA sequence; (h) having a 16S rDNA sequence identical to the sequence of SEQ ID NO:6; (i) having a 16S rDNA sequence identical to the sequence of SEQ ID NO:7; (j) a SEQ ID NO:8 A 16S rDNA sequence that is identical to the sequence of SEQ ID NO:; (k) has a 16S rDNA sequence that is identical to the sequence of SEQ ID NO:9; and/or (l) is a 16S rDNA sequence that is identical to the sequence of SEQ ID NO:10 Have.

SEQ ID NO: 5 is as follows (mutations compared to the 16SA sequence of the reference strain KPA171202 are underlined).

SEQ ID NO: 6 is as follows (mutations compared to the 16S sequence of the reference strain KPA171202 are underlined).

SEQ ID NO:7 is as follows (mutations compared to the 16S sequence of the reference strain KPA171202 are underlined).

SEQ ID NO: 8 is as follows (mutations compared to the 16S sequence of the reference strain KPA171202 are underlined).

SEQ ID NO: 9 is as follows (mutations compared to the 16S sequence of the reference strain KPA171202 are underlined).

SEQ ID NO: 10 is as follows (mutations compared to the 16S sequence of the reference strain KPA171202 are underlined).

In embodiments, the at least one additional bacterium comprises, consists essentially of, or consists of probiotic bacteria. In embodiments, the at least one bacterium is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 bacterial strains and/or strains, about 10, 9, 8, 7, 6, Less than 5, 4, 3 or 2 bacterial strains and/or bacterial species, or 1-10, 2-10, 3-10, 4-10, 5-10, 1-5, 2-5, 3-5 or Includes 4-5 bacterial strains and/or strains. In embodiments, the at least one bacterium comprises a plurality of bacterial strains and/or bacterial species, eg, at least about 2,3,4,5,6,7,8,9,10 bacterial strains and/or bacterial species. Including. In an embodiment, the at least one bacterium is an isolated Propionibacterium granulosum bacterium, an isolated Propionibacterium avidum bacterium, an isolated Staphylococcus epidermidis bacterium, a simple bacterium. It includes isolated Staphylococcus aureus and/or isolated Corynebacterium jeikeium. In an embodiment, the at least one bacterium is an isolated Propionibacterium granulosum, an isolated Propionibacterium avidum, an isolated Staphylococcus epidermides, an isolated Staphylococcus epidermides. One, two (any combination thereof), three (any combination thereof), four (any combination of) of the phylococcus aureus and/or the isolated Corynebacterium jaykeiham ) Or 5 are included.

In embodiments, the compositions or combinations provided herein include an enhancing peptide or enzyme. In embodiments, the enhancing peptide or enzyme has the following properties: biofilm degradation, improved skin penetration, antibacterial, inflammation (eg, skin) relief, hypersensitivity (eg, skin) relief, redness (eg, One or more or any combination of: (skin) relief, skin tightening, wrinkle removal, fine wrinkle removal or other appearance (eg, skin) improvement.

In an embodiment, P. A composition comprising an acnes bacteriophage and an anti-acne compound is provided. In embodiments, the composition comprises a pharmaceutically acceptable carrier. In an embodiment, P. The dose of acnes bacteriophage is adjusted (eg, increased or decreased) for stability. In an embodiment, P. The dose of acnes bacteriophage is adjusted up or down depending on the anti-acne compound due to its stability in combination with the anti-acne compound.

In an embodiment, P. A combination or system is provided that comprises an acnes bacteriophage and one or more anti-acne compounds. In an example, the bacteriophage are in one composition (eg, in one container, such as a bottle, tube, or other container), and one or more anti-acne compounds are in another composition (eg, , In another container, such as a bottle, tube, or other container). In embodiments, the bacteriophage-containing composition comprises a pharmaceutically acceptable carrier. In embodiments, the composition comprising the anti-acne compound comprises a pharmaceutically acceptable carrier. In embodiments, the additional one or more compounds (eg, enzymes, hydrating compounds, UV absorbing or blocking compounds, etc.) is a composition comprising a bacteriophage, a composition comprising one or more anti-acne compounds, or 3 in another composition (eg, in a third container, such as a bottle, tube or other container). In an embodiment, the one or more probiotic bacteria is in a composition comprising a bacteriophage, a composition comprising one or more anti-acne compounds or a third separate composition (eg, bottle, tube or Present in a third container, like the other containers). In embodiments, the combination or system further comprises instructions for administration. In embodiments, the combination or system has at least about 2, 3, 4, 5, 6, 7, 8, 9 or 10 P. Includes acnes bacteriophage.

In an embodiment, P. A combination or system is provided that comprises an Acnes bacteriophage and one or more probiotic bacteria and/or one or more compounds (such as one or more enzymes or anti-acne compounds). In an example, bacteriophage are in one composition (eg, in one container such as a bottle, tube or other container) and one or more probiotic bacteria are in another composition (in a bottle). , In a separate container, such as a tube or other container), optionally wherein the additional one or more compounds comprises a composition comprising a bacteriophage, a composition comprising one or more probiotic bacteria. Or in a third separate composition (in a third container such as a bottle, tube or other container). In embodiments, the combination or system further comprises instructions for administration. In embodiments, the combination or system has at least about 2, 3, 4, 5, 6, 7, 8, 9 or 10 P. Includes acnes bacteriophage.

In embodiments, the system, combination or composition comprises an enzyme such as a biofilm degrading enzyme or an anti-aging enzyme. Non-limiting examples of biofilm degrading enzymes are deoxyribonucleases (eg deoxyribonuclease I), proteases (eg papain, bromelain, trypsin, proteinase K, subtilisin or seratiopeptidase), glycosidases (eg dispascin, alginate lyase). , Amylase or cellulase). Non-limiting examples of anti-aging enzymes include superoxide dismutase and peroxidase.

In embodiments, the system, combination or composition comprises a topical retinoid, an antibiotic and/or an alpha hydroxy acid. In embodiments, the system or composition further comprises a topical retinoid. In embodiments, the system or composition further comprises an antibiotic. In embodiments, the system or composition further comprises an alpha hydroxy acid. In embodiments, the system or composition further comprises benzoyl peroxide, salicylic acid, sulfur, resorcinol, resorcinol monoacetate or any combination thereof. In embodiments, benzoyl peroxide is 2.5% to 10%, such as about 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%. %, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5% or 10% (weight/volume). In embodiments, benzoyl peroxide is present at a concentration of less than 2.5% but greater than about 0.1%, 0.5%, 1%, 1.5% or 2% (weight/volume). ing. In embodiments, salicylic acid is 0.5% to 2%, such as about 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.1%. , 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9% or 2% (weight/volume) concentration Exists in. In embodiments, salicylic acid is present at a concentration of less than 0.5% but greater than about 0.1% (weight/volume). In embodiments, sulfur is 3% to 10%, such as about 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%. , 7.5%, 8%, 8.5%, 9%, 9.5% or 10% (weight/volume). In embodiments, sulfur is less than 3% but is present at a concentration greater than about 0.1%, 0.5%, 1%, 1.5%, 2% or 2.5% (weight/volume). doing. In embodiments, resorcinol is present at a concentration of 2% and sulfur is between 3% and 8% (eg, about 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5%, 0.5%, 6%, 6.5%, 7%, 7.5% or 8%) (weight/volume). In an embodiment, resorcinol monoacetate is present at a concentration of 3% and sulfur is between 3% and 8% (eg, about 2.5%, 3%, 3.5%, 4%, 4.5%, 5%. 5.5%, 6%, 6.5%, 7%, 7.5% or 8%) (weight/volume). In embodiments, resorcinol is present at a concentration of less than 2% but greater than about 0.1%, 0.5%, 1%, 1.5% (weight/volume). In embodiments, the resorcinol monoacetate has a concentration of less than 3% but greater than about 0.1%, 0.5%, 1%, 1.5%, 2% or 2.5% (weight/volume). Exists in.

In embodiments, the compositions provided herein include a moisturizer.

Methods of Treating Acne In an aspect, provided herein is a method of preventing or treating acne in a subject in need thereof, the method comprising: Administering an effective amount of a composition or combination provided herein. In an embodiment, at least one P. An effective amount of a composition comprising, consisting essentially of, or consisting of an acnes bacteriophage, at least one anti-acne compound and a pharmaceutically acceptable carrier is administered to the subject. In an embodiment, at least one P. A probiotic bacterium-free composition is administered to a subject in an effective amount of a composition comprising an acnes bacteriophage, at least one anti-acne compound, and a pharmaceutically acceptable carrier. In an embodiment, P. An effective amount of a composition comprising an acnes bacteriophage and an enzyme is administered to the subject.

In embodiments, an effective amount of a bacteriophage-comprising composition described herein, including this embodiment, is administered to the subject. In an embodiment, the bacteriophage is a wild type bacteriophage.

In embodiments, the bacteriophage is administered locally. In embodiments, the bacteriophage is in a composition (eg, a pharmaceutical or cosmetic composition) that further comprises a pharmaceutically or cosmetically acceptable carrier.

In embodiments, the method further comprises administering probiotic bacteria to the subject.

In an aspect, there is provided a method of treating acne in a subject in need thereof. The method is described by Probiotic P.P. Administering to the subject an effective amount of Acnes. In embodiments, the method further comprises administering a bacteriophage to the subject.

In an embodiment, P. Acnes has a 16S rDNA sequence containing the T992C, T838C, C1322T and/or C986T mutations compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2. In an embodiment, P. Acnes contains a 16S rDNA sequence with the T838C and C1322T mutations as compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2. In an embodiment, P. Acnes is ProI strain. In an embodiment, P. Acnes contains a 16S rDNA sequence with the C986T and T992C mutations as compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2. In an embodiment, P. Acnes is a ProII strain.

In an embodiment, P. Acnes contains (a) a 16S rDNA sequence with the T992C mutation as compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO: 2; (b) a KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO: 2 In comparison, it contains a 16S rDNA sequence with the T838C mutation; (c) contains a 16S rDNA sequence with the C1322T mutation as compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2; (d) SEQ ID NO:2 Comprising a 16S rDNA sequence with a C986T mutation, as compared to the KPA171202 reference strain 16S rDNA sequence shown as: (e) containing a 16S rDNA sequence identical to the sequence of SEQ ID NO:3; (g) a sequence of SEQ ID NO:4 A 16S rDNA sequence that is identical to:; (h) no linear plasmid; (i) no plasmid containing the virulence factor; and/or (j) extrachromosomal lipase and/or adherent virulence factor It does not include a plasmid encoding

In embodiments, the method further comprises administering to the subject at least one additional probiotic bacterium.

In embodiments, the at least one additional bacterium comprises, consists essentially of, or consists of probiotic bacteria. In embodiments, the at least one bacterium is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 bacterial strains and/or strains, about 10, 9, 8, 7, 6, Less than 5, 4, 3 or 2 bacterial strains and/or bacterial species, or 1-10, 2-10, 3-10, 4-10, 5-10, 1-5, 2-5, 3-5 or Includes 4-5 bacterial strains and/or strains. In embodiments, the at least one bacterium comprises a plurality of bacterial strains and/or bacterial species, eg, at least about 2,3,4,5,6,7,8,9,10 bacterial strains and/or bacterial species. Including. In embodiments, the at least one bacterium comprises a Propionibacterium sp., Staphylococcus sp. and/or Corynebacterium sp. In embodiments, the at least one bacterium comprises a bacterium within the class Beta Proteobacteria. In an embodiment, the at least one bacterium is an isolated Propionibacterium granulosum, an isolated Propionibacterium avidum, an isolated Staphylococcus epidermides, an isolated Staphylococcus epidermides. Includes Phylococcus aureus and/or isolated Corynebacterium jaykeiham. In an embodiment, the at least one bacterium is an isolated Propionibacterium granulosum, an isolated Propionibacterium avidum, an isolated Staphylococcus epidermides, an isolated Staphylococcus epidermides. Includes one, two, three, four or five of the phylococcus aureus and/or the isolated Corynebacterium jaykeiham.

In an embodiment, the subject has been administered the bacteriophage described herein, including this embodiment.

In an embodiment, the subject is P. Has been given antibiotics to kill Acnes. In embodiments, the antibiotic is clindamycin, doxycycline, erythromycin or tetracycline, or a derivative of clindamycin, doxycycline, erythromycin or tetracycline.

In embodiments, the antibiotic is clindamycin, doxycycline, erythromycin or tetracycline, or a derivative of clindamycin, doxycycline, erythromycin or tetracycline.

In embodiments, the method further comprises administering to the subject an enzyme such as a biofilm degrading enzyme or an anti-aging enzyme. Non-limiting examples of biofilm degrading enzymes include deoxyribonucleases (eg deoxyribonuclease I), restriction endonucleases, proteases (eg papain, bromelain, trypsin, proteinase K, subtilisin or seratiopeptidase), glycosidases (eg dispase). Syn, alginate lyase, amylase or cellulase). Non-limiting examples of anti-aging enzymes include superoxide dismutase and peroxidase.

In embodiments, the method further comprises administering a topical retinoid, an antibiotic and/or an alpha hydroxy acid. In embodiments, the method further comprises administering a topical retinoid. In embodiments, the method further comprises administering an antibiotic. In embodiments, the method further comprises administering an alpha hydroxy acid. In embodiments, the method further comprises administering benzoyl peroxide, salicylic acid, sulfur, resorcinol and/or resorcinol monoacetate to the subject. In embodiments, the method further comprises administering benzoyl peroxide. In embodiments, the method further comprises administering salicylic acid. In embodiments, the method further comprises administering sulfur. In embodiments, the method further comprises administering resorcinol and/or sulfur. In embodiments, the method further comprises administering resorcinol and/or resorcinol monoacetate.

In embodiments, the method further comprises administering the enhancing peptide or enzyme. In embodiments, the enhancing peptide or enzyme has the following properties: biofilm degradation, improved skin penetration, antibacterial, relief of inflammation (eg, skin), relief of hypersensitivity (eg, of skin), redness (eg, of skin). One or more or any combination of: (skin) relief, skin tightening, wrinkle removal, fine wrinkle removal or other appearance (eg, skin) improvement.

In embodiments, at least about 2, 3, 4, 5, 6, 7, 8, 9 or 10 P. An acnes bacteriophage is administered to the subject. In an embodiment, P. Acnes bacteriophage is more than one type of P. Includes acnes bacteriophage.

Exemplary Methods and Compositions for Treating Acne In embodiments, compositions comprising bacteriophage are provided herein. In embodiments, the bacteriophage is present in a composition such as a therapeutic or cosmetic composition. In embodiments, the composition further comprises a strain of probiotic bacteria. In embodiments, the composition further comprises an enzyme that degrades bacterial biofilm (eg, its constituents) at or on human skin pores. In embodiments, the enzyme enhances penetration of bacteriophage and/or probiotic bacteria. In an embodiment, the bacteriophage (“phage”) does not kill the beneficial skin bacteria, but the P. It destroys acne acne-causing (ie, pathogenic) strains with a high degree of specificity and effectiveness. In embodiments, the biofilm degrading enzyme lyses the biofilm to sensitize the pathogen (eg, by reducing pathogen adhesion to the host cell and/or increasing bacteriophage access to the pathogenic cell. By that). In embodiments, the probiotic bacterium is immune to the bacteriophage (eg, the bacterium lacks a cell receptor to which the bacteriophage specifically binds). In an embodiment, the probiotic bacterium is sterilized P. Acnes occupy the niche left by pathogenic strains. In embodiments, the probiotic bacterium reduces or prevents recolonization or growth on the subject's skin (such as pores) by surviving longer than the pathogenic bacterium.

In an aspect, a composition is provided for the therapeutic treatment of the skin disease acne. In an embodiment, the composition is a lytic P. Acnes bacteriophage, optionally containing a probiotic bacterium of healthy skin origin and/or optionally a biomembrane degrading enzyme in the composition as an adjuvant to enhance penetration of the active component ..

In an embodiment, a lytic P. Acnes bacteriophage is a pathogenic P. aeruginosa on skin comedones. Get infected with acnes. In an embodiment, the bacteriophage is P. Duplicate and dissolve it in Acnes. In an embodiment, P. When Acnes dissolves, it releases new virions. In an embodiment, the enzyme clears the blockage of the blocked comedone, P. Acnes acnes biofilm is dissolved and P. Increases the virion's approach to Acnes. In an embodiment, a lytic P. The exponential growth of Acnes phages, without interfering with the growth of beneficial skin symbiotic bacteria, has led to P. Rapidly sterilizes Acnes with high specificity. In an embodiment, P. The niche vacated by Acnes is then filled with probiotic bacteria. In an embodiment, the bacteria originate from healthy skin and spread to occupy a niche, thereby causing any surviving P. cerevisiae. Prevent the Acnes bacteria from growing and returning to its original state. In embodiments, this strategy helps balance the skin microbiome in the subject and reconditions the microbiome towards a healthy skin bacterial population. In embodiments, the biofilm degrading enzyme is in the formulation as an adjuvant that helps clear blockages of the blocked comedones and increases access of phage and probiotic bacteria to the pores.

In an embodiment, a combination is provided that includes a bacteriophage, a probiotic bacterium and an enzyme that (optionally) enhances penetration of the bacteriophage. In an embodiment, the pathogen is killed and the probiotic bacterium replaces the pathogen. In embodiments, a "sterilize and replace" approach is used to treat acne. In an embodiment, a subject is administered a biologic that selectively kills pathogenic bacteria that cause acne. In embodiments, healthy skin-sourced probiotic bacteria occupy a niche of applied and killed pathogens. In embodiments, this approach avoids the problem of uncontrollable drug resistance associated with antibiotics. In embodiments, the presence of actively dividing probiotic bacteria prevents recurrence by preventing any pathogen from growing and returning. In embodiments, dysbiosis on the skin of the subject is treated. In embodiments, the microbiome associated with acne is readjusted into a healthy microbiome.

In an embodiment, the bacteriophage is a naturally-occurring P. Acnes bacteriophage.

Non-limiting examples of enzymes that may be co-administered with bacteriophage include BL00275 from Bacillus licheniformis; deoxyribonuclease I; restriction endonuclease, deoxyribonuclease (eg Staphylococcus aureus thermonuclease, B Licheniformis NucB, derived from deoxyribonuclease 1L2); glycoside hydrolases (eg, dispersin B, alginate lyase, amylase, cellulase, glycanase); and proteases (eg, subtilisin, proteinase K, trypsin, seratiopeptidase).

Non-limiting examples of probiotic bacteria that may be administered or may be present in the system or composition include the following bacterial species: Propionibacterium acnes, Propionibacterium granulosum, Includes one or more or any combination of Propionibacterium avidum, Staphylococcus epidermides, Staphylococcus aureus, and Corynebacterium jaykeiam. In an embodiment, the probiotic bacterial strain is characterized by (a) low lipase activity and low adhesion to human keratinocytes, and colonizes the skin without inducing a deleterious immune response; and (b) propioni Selected based on its ability to occupy a niche similar to Bacterium acnes.

In embodiments, a biofilm degrading enzyme is present in the formulation and acts as an adjuvant that enhances the effectiveness of the active ingredient (such as bacteriophage). In an embodiment, the enzyme is P. It has the ability to degrade the Acnes biofilm in vitro.

Embodiments Embodiments and examples are provided below to facilitate a more complete understanding of the present invention. The following embodiments and examples describe exemplary ways of making and carrying out the present invention. However, the scope of the present invention is not limited to the particular embodiments disclosed in these embodiments and examples, as these embodiments and examples provide similar results if alternative methods are utilized. , For the purpose of explanation only.

The embodiments include the following embodiments P1 to P56.

Embodiment P1. A composition consisting essentially of at least one Propionibacterium acnes bacteriophage, at least one anti-acne compound and a pharmaceutically acceptable carrier.

Embodiment P2. A composition comprising at least one Propionibacterium acnes bacteriophage, at least one anti-acne compound and a pharmaceutically acceptable carrier and free of probiotic bacteria.

Embodiment P3. The composition is P. The composition of Embodiment P2 further comprising an Acnes biofilm degrading enzyme.

Embodiment P4. The composition of any one of Embodiments P1-P3 wherein the at least one anti-acne compound is benzoyl peroxide.

Embodiment P5. The composition of Embodiment P4 wherein benzoyl peroxide is present at a concentration of 2.5%-10% (weight/volume).

Embodiment P6. Benzoyl peroxide is present at a concentration of less than 2.5% but greater than about 0.1%, 0.5%, 1%, 1.5% or 2% (weight/volume). A composition of form P4.

Embodiment P7. The composition of any one of Embodiments P1 through P3 wherein the at least one anti-acne compound is salicylic acid.

Embodiment P8. The composition of Embodiment P7 wherein salicylic acid is present at a concentration of 0.5% to 2% (weight/volume).

Embodiment P9. The composition of Embodiment P7 wherein salicylic acid is present in a concentration of less than 0.5% but greater than about 0.1% (weight/volume).

Embodiment P10. The composition of any one of Embodiments P1 through P3 wherein the at least one anti-acne compound is sulfur.

Embodiment P11. The composition of Embodiment P10 wherein sulfur is present in a concentration from 3% to 10% (weight/volume).

Embodiment P12. Sulfur is present in a concentration of less than 3% but greater than about 0.1%, 0.5%, 1%, 1.5%, 2% or 2.5% (weight/volume), The composition of Embodiment P10.

Embodiment P13. The composition of any one of Embodiments P1 through P3 wherein the at least one anti-acne compound is resorcinol and sulfur.

Embodiment P14. The composition of Embodiment P13 wherein resorcinol is present in a concentration of 2% and sulfur is present in a concentration of 3% to 8% (weight/volume).

Embodiment P15. The composition of any one of Embodiments P1-P3 wherein the at least one anti-acne compound comprises resorcinol monoacetate and sulfur.

Embodiment P16. The composition of Embodiment P15 wherein resorcinol monoacetate is present at a concentration of 3% and sulfur is present at a concentration of 3% to 8% (weight/volume).

Embodiment P17. The composition of any one of Embodiments P1 through P3 wherein the anti-acne compound is an antibiotic, retinoid or alpha hydroxy acid.

Embodiment P18. A composition comprising Propionibacterium acnes bacteriophage and an enzyme.

Embodiment P19. P. Acnes bacteriophage is a lytic P. The composition of any one of Embodiments P1 through P18 which is an Acnes bacteriophage.

Embodiment P20. P. The composition of any one of embodiments P1-P19, wherein the Acnes bacteriophage comprises a linear double-stranded DNA genome.

Embodiment P21. P. The composition of any one of Embodiments P1-P20, wherein the Acnes bacteriophage is within the family of Bacteriophage Siphoviruses.

Embodiment P22. P. Embodiment P1 wherein the Acnes bacteriophage genome comprises a nucleotide sequence that is at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to the nucleotide sequence of SEQ ID NO:1. ~ The composition of any one of P21.

Embodiment P23. The enzyme is P. The composition of any one of embodiments P18-P21, which is an Acnes biofilm degrading enzyme.

Embodiment P24. The composition of any one of Embodiments P3 or P18-P23, wherein the enzyme is a glycosidase, protease, deoxyribonuclease or restriction endonuclease.

Embodiment P25. The composition of any one of Embodiments P3 or P18-P24 wherein the enzyme is a glycosidase.

Embodiment P26. The composition of Embodiment P25 wherein the glycosidase is a glycoside hydrolase.

Embodiment P27. The composition of Embodiment P26 wherein the enzyme catalyzes the hydrolysis of a linear polymer of N-acetyl-D-glucosamine.

Embodiment P28. The composition of Embodiment P27 wherein the enzyme is β-hexosaminidase.

Embodiment P29. The composition of Embodiment P28 wherein the enzyme hydrolyzes the β-1,6-glycosidic bond of the acetylglucosamine polymer.

Embodiment P30. The enzyme is deoxyribonuclease I, a restriction endonuclease, papain, bromelain, trypsin, proteinase K, subtilisin, seratiopeptidase, dispasin, alginate lyase, amylase or cellulase, or any one of embodiments P3 or P18 to P24. Composition.

Embodiment P31. The composition of any one of Embodiments P3 or P18-P24 wherein the enzyme is Dispersin B.

Embodiment P32. The composition of any one of Embodiments P3 or P18-P24, wherein the enzyme is a protease and the protease is proteinase K or subtilisin.

Embodiment P33. The composition of any one of embodiments P18-P22, wherein the enzyme is an anti-aging enzyme.

Embodiment P34. The composition of Embodiment P33 wherein the anti-aging enzyme is superoxide dismutase or peroxidase.

Embodiment P35. The composition of any one of Embodiments P18-P34 further comprising probiotic bacteria.

Embodiment P36. The probiotic bacteria are probiotic P. The composition of Embodiment P35 which is a genus species, Staphylococcus species and/or Corynebacterium species bacteria.

Embodiment P37. The composition of Embodiment P35 wherein the probiotic bacterium is a bacterium within the class of Beta Proteobacteria.

Embodiment P38. The probiotic bacteria are probiotic P. The composition of Embodiment P36 which is an Acnes bacterium.

Embodiment P39. P. Acnes bacteria
(A) comprises a 16S DNA sequence with a T992C mutation as compared to the KPA171202 reference strain 16S DNA sequence shown as SEQ ID NO:2;
(B) comprises a 16S DNA sequence with a T838C mutation as compared to the KPA171202 reference strain 16S DNA sequence shown as SEQ ID NO:2;
(C) contains a 16S DNA sequence with the C1322T mutation as compared to the KPA171202 reference strain 16S DNA sequence shown as SEQ ID NO:2;
(D) contains a 16S DNA sequence with a C986T mutation as compared to the KPA171202 reference strain 16S DNA sequence shown as SEQ ID NO:2;
(E) comprises a 16S DNA sequence that is identical to the sequence of SEQ ID NO:3;
(F) comprises a 16S DNA sequence that is identical to the sequence of SEQ ID NO:4;
(G) does not contain a linear plasmid;
(H) does not contain a plasmid containing a virulence factor; and/or (i) does not contain a plasmid encoding an extrachromosomal lipase and/or an adherent virulence factor,
The composition of Embodiment P38.

Embodiment P40. P. Acnes bacteria
(A) Pathogenic P. cerevisiae when grown in suspension culture. Produces less than about 20% of the level of lipase produced by the Acnes strain;
(B) Pathogenic P. cerevisiae when grown in adherent culture. Produces less than about 10% of the level of lipase produced by the Acnes strain;
(C) Adhesion to epithelial cells causes pathogenic P. At least 50% less than the Acnes strain, and/or (d) pathogenic P. Less inflammatory than Acnes strain,
The composition of Embodiment P38.

Embodiment P41. The composition of any one of embodiments P35-P40, further comprising at least one additional probiotic bacterium.

Embodiment P42. The composition of Embodiment P41 wherein the at least one additional probiotic bacterium comprises Propionibacterium granulosum and/or Propionibacterium avidum.

Embodiment P43. The pathogenic P. Acnes strains,
(A) comprises a 16S DNA sequence with a G1058C mutation as compared to the KPA171202 reference strain 16S DNA sequence shown as SEQ ID NO:2;
(B) contains the 16S DNA sequence with the G1058C and A1201C mutations compared to the KPA171202 reference strain 16S DNA sequence shown as SEQ ID NO:2;
(C) contains a 16S DNA sequence with a G529A mutation as compared to the KPA171202 reference strain 16S DNA sequence shown as SEQ ID NO:2;
(D) contains the 16S DNA sequence with the G1004A and T1007C mutations compared to the KPA171202 reference strain 16S DNA sequence shown as SEQ ID NO:2;
(E) comprises a 16S DNA sequence with the G1268A mutation as compared to the KPA171202 reference strain 16S DNA sequence shown as SEQ ID NO:2;
(F) comprises a 16S DNA sequence with the T554C and G1058C mutations as compared to the KPA171202 reference strain 16S DNA sequence shown as SEQ ID NO:2;
(G) comprises a 16S DNA sequence which is identical to the sequence of SEQ ID NO:5;
(H) comprises a 16S DNA sequence that is identical to the sequence of SEQ ID NO:6;
(I) comprises a 16S DNA sequence that is identical to the sequence of SEQ ID NO:7;
(J) contains a 16S DNA sequence that is identical to the sequence of SEQ ID NO:8;
(K) comprises a 16S DNA sequence identical to the sequence of SEQ ID NO: 9; and/or (l) comprises a 16S DNA sequence identical to the sequence of SEQ ID NO: 10,
The composition of Embodiment P40.

Embodiment P44. At least one additional P. The composition of any one of Embodiments P18-P43 further comprising an Acnes bacteriophage.

Embodiment P45. The composition of any one of Embodiments P1-P44 comprising a pharmaceutically acceptable carrier.

Embodiment P46. The composition of Embodiment P45 wherein the pharmaceutically acceptable carrier comprises an emulsion.

Embodiment P47. The composition of Embodiment P46 wherein the emulsion is an oil-in-water emulsion or a water-in-oil emulsion.

Embodiment P48. The composition of any one of Embodiments P1-P47, which is in the form of a cream, lotion, suspension or aqueous solution.

Embodiment P49. A combination consisting essentially of at least one Propionibacterium acnes bacteriophage and at least one anti-acne compound, each of at least one Propionibacterium acnes bacteriophage and at least one anti-acne compound Wherein the combination is in a composition further comprising a pharmaceutically acceptable carrier.

Embodiment P50. At least one P. The combination of Embodiment P49 wherein the Acnes bacteriophage and the at least one anti-acne compound are in separate compositions.

Embodiment P51. At least one P. The combination of Embodiment P50 wherein the Acnes bacteriophage and the at least one anti-acne compound are in separate containers.

Embodiment P52. A combination comprising Propionibacterium acnes bacteriophage and an enzyme.

Embodiment P53. P. The combination of embodiment P52 wherein the acnes bacteriophage and enzyme are in separate compositions.

Embodiment P54. P. The combination of embodiment P53 wherein the Acnes bacteriophage and enzyme are in separate containers.

Embodiment P55. A method of treating acne in a subject in need thereof for treating an effective amount of a composition of any one of embodiments P1-P46 or a combination of any one of embodiments P49-P54. A method comprising administering.

Embodiment P56. The method of embodiment P55 wherein the composition is administered topically.

Additional embodiments include the following embodiments 1-55.

Embodiment 1. A composition comprising at least one Propionibacterium acnes bacteriophage, at least one anti-acne compound and a pharmaceutically acceptable carrier.

Embodiment 2. The composition of Embodiment 1 free of probiotic bacteria.

Embodiment 3. The composition is P. The composition of Embodiment 1 or 2, further comprising an Acnes biofilm degrading enzyme.

Embodiment 4. The composition of any one of embodiments 1-3, wherein the at least one anti-acne compound is salicylic acid.

Embodiment 5. The composition of Embodiment 4 wherein the salicylic acid is present at a concentration of 0.5% to 2% (weight/volume).

Embodiment 6. The composition of Embodiment 5 wherein the salicylic acid is present at a concentration of less than 0.5% but greater than about 0.1% (weight/volume).

Embodiment 7. The composition of any one of embodiments 1-3, wherein the at least one anti-acne compound is sulfur.

Embodiment 8. The composition of Embodiment 7 wherein sulfur is present in a concentration of 3% to 10% (weight/volume).

Embodiment 9. Sulfur is present in a concentration of less than 3% but greater than about 0.1%, 0.5%, 1%, 1.5%, 2% or 2.5% (weight/volume), The composition of Embodiment 7.

Embodiment 10. The composition of any one of embodiments 1-3, wherein the at least one anti-acne compound is resorcinol and sulfur.

Embodiment 11. The composition of Embodiment 10 wherein resorcinol is present in a concentration of 2% and sulfur is present in a concentration of 3% to 8% (weight/volume).

Embodiment 12. The composition of any one of embodiments 1-3, wherein the at least one anti-acne compound comprises resorcinol monoacetate and sulfur.

Embodiment 13. The composition of Embodiment 12 wherein resorcinol monoacetate is present at a concentration of 3% and sulfur is present at a concentration of 3% to 8% (weight/volume).

Embodiment 14. The composition of any one of embodiments 1-3, wherein the anti-acne compound is an antibiotic, a retinoid or an alpha hydroxy acid.

Embodiment 15. The composition of any one of embodiments 1-14, wherein the Propionibacterium acnes bacteriophage is a naturally occurring Propionibacterium acnes bacteriophage.

Embodiment 16. P. Acnes bacteriophage is a lytic P. The composition of any one of embodiments 1-15, which is an acnes bacteriophage.

Embodiment 17. P. The composition of any one of embodiments 1-16, wherein the Acnes bacteriophage comprises a linear double stranded DNA genome.

Embodiment 18. P. The composition of any one of embodiments 1-17, wherein the acnes bacteriophage is within the family of bacteriophage siphovirus.

Embodiment 19. P. Embodiment 1 wherein the Acnes bacteriophage genome comprises a nucleotide sequence that is at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to the nucleotide sequence of SEQ ID NO:1. -19. The composition of any one of -19.

Embodiment 20. The enzyme is P. The composition of any one of embodiments 3-19, which is an Acnes biofilm degrading enzyme.

Embodiment 21. The composition of any one of embodiments 3-20, wherein the enzyme is a glycosidase, protease, deoxyribonuclease or restriction endonuclease.

Embodiment 22. 22. The composition of any one of embodiments 3-21, wherein the enzyme is a glycosidase.

Embodiment 23. The composition of Embodiment 22 wherein the glycosidase is a glycoside hydrolase.

Embodiment 24. The composition of Embodiment 23 wherein the enzyme catalyzes the hydrolysis of a linear polymer of N-acetyl-D-glucosamine.

Embodiment 25. The composition of Embodiment 24 wherein the enzyme is β-hexosaminidase.

Embodiment 26. The composition of Embodiment 25 wherein the enzyme hydrolyzes the β-1,6-glycosidic bond of the acetylglucosamine polymer.

Embodiment 27. The composition of any one of embodiments 3-20, wherein the enzyme is deoxyribonuclease I, a restriction endonuclease, papain, bromelain, trypsin, proteinase K, subtilisin, seratiopeptidase, dispasin, alginate lyase, amylase or cellulase. .

Embodiment 28. The composition of any one of embodiments 3-20, wherein the enzyme is Dispersin B.

Embodiment 29. The composition of any one of embodiments 3-20, wherein the enzyme is a protease and the protease is proteinase K or subtilisin.

Embodiment 30. The composition of any one of embodiments 1-29, further comprising an anti-aging enzyme.

Embodiment 31. The composition of Embodiment 30 wherein the anti-aging enzyme is superoxide dismutase or peroxidase.

Embodiment 32. The composition of any one of embodiments 1-31, further comprising probiotic bacteria.

Embodiment 33. The probiotic bacteria are probiotic P. The composition of Embodiment 32, which is a genus species, Staphylococcus species and/or Corynebacterium species bacteria.

Embodiment 34. The composition of Embodiment 32 wherein the probiotic bacterium is a bacterium within the class of beta-proteobacteria.

Embodiment 35. The probiotic bacteria are probiotic P. The composition of Embodiment 33 which is an Acnes bacterium.

Embodiment 36. P. Acnes bacteria
(A) contains the 16S rDNA sequence with the T992C mutation as compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2;
(B) contains the 16S rDNA sequence with the T838C mutation as compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2;
(C) contains a 16S rDNA sequence with the C1322T mutation as compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2;
(D) contains the 16S rDNA sequence with the C986T mutation as compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2;
(E) comprises a 16S rDNA sequence that is identical to the sequence of SEQ ID NO:3;
(F) comprises a 16S rDNA sequence that is identical to the sequence of SEQ ID NO:4;
(G) does not contain a linear plasmid;
(H) does not contain a plasmid containing a virulence factor; and/or (i) does not contain a plasmid encoding an extrachromosomal lipase and/or an adherent virulence factor,
The composition of Embodiment 35.

Embodiment 37. P. Acnes bacteria
(A) Pathogenic P. cerevisiae when grown in suspension culture. Produces less than about 20% of the level of lipase produced by the Acnes strain;
(B) Pathogenic P. cerevisiae when grown in adherent culture. Produces less than about 10% of the level of lipase produced by the Acnes strain;
(C) Adhesion to epithelial cells causes pathogenic P. At least 50% less than the Acnes strain, and/or (d) pathogenic P. Less inflammatory than Acnes strain,
The composition of Embodiment 35 or 36.

Embodiment 38. The composition of any one of embodiments 32-37, further comprising at least one additional probiotic bacterium.

Embodiment 39. 39. The composition of embodiment 38, wherein the at least one additional probiotic bacterium comprises Propionibacterium granulosum and/or Propionibacterium avidum.

Embodiment 40. The pathogenic P. Acnes strains,
(A) contains the 16S rDNA sequence with the G1058C mutation as compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2;
(B) containing the 16S rDNA sequence with the G1058C and A1201C mutations compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2;
(C) contains the 16S rDNA sequence with the G529A mutation as compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2;
(D) contains the 16S rDNA sequence with the G1004A and T1007C mutations compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2;
(E) contains the 16S rDNA sequence with the G1268A mutation as compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2;
(F) contains the 16S rDNA sequence with the T554C and G1058C mutations compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2;
(G) contains a 16S rDNA sequence that is identical to the sequence of SEQ ID NO:5;
(H) comprises a 16S rDNA sequence that is identical to the sequence of SEQ ID NO:6;
(I) comprises a 16S rDNA sequence that is identical to the sequence of SEQ ID NO:7;
(J) comprises a 16S rDNA sequence which is identical to the sequence of SEQ ID NO:8;
(K) comprising a 16S rDNA sequence identical to the sequence of SEQ ID NO: 9; and/or (l) comprising a 16S rDNA sequence identical to the sequence of SEQ ID NO: 10,
The composition of Embodiment 37.

Embodiment 41. At least one additional P. The composition of any one of embodiments 1-40, further comprising an acnes bacteriophage.

Embodiment 42. 42. The composition of any one of embodiments 1-41, wherein the pharmaceutically acceptable carrier comprises an emulsion.

Embodiment 43. The composition of Embodiment 42 wherein the emulsion is an oil-in-water emulsion or a water-in-oil emulsion.

Embodiment 44. The composition of any one of embodiments 1-44, which is in the form of a cream, lotion, suspension or aqueous solution.

Embodiment 45. A combination comprising at least one Propionibacterium acnes bacteriophage and at least one anti-acne compound, wherein each of the at least one Propionibacterium acnes bacteriophage and at least one anti-acne compound is a pharmaceutical A combination in the composition further comprising a pharmaceutically acceptable carrier.

Embodiment 46. At least one P. The combination of embodiment 45 wherein the acnes bacteriophage and the at least one anti-acne compound are in separate compositions.

Embodiment 47. The combination of embodiment 46 wherein the at least one anti-acne compound is benzoyl peroxide.

Embodiment 48. The combination of embodiment 47 wherein benzoyl peroxide is present at a concentration of 2.5%-10% (weight/volume).

Embodiment 49. An embodiment in which benzoyl peroxide is less than 2.5% but is present in a concentration greater than about 0.1%, 0.5%, 1%, 1.5% or 2% (weight/volume). 47 combinations.

Embodiment 50. 47. A method of treating acne in a subject in need thereof, comprising administering to the subject an effective amount of the composition of any one of embodiments 1-44.

Embodiment 51. The method of embodiment 50 wherein the composition is administered topically.

Embodiment 52. 50. A method of treating acne in a subject in need thereof, comprising administering to the subject an effective amount of the combination of any one of embodiments 45-49.

Embodiment 53. A composition comprising Propionibacterium acnes bacteriophage and an enzyme.

Embodiment 54. A combination comprising Propionibacterium acnes bacteriophage and an enzyme.

Embodiment 55. A composition consisting essentially of at least one Propionibacterium acnes bacteriophage, at least one anti-acne compound and a pharmaceutically acceptable carrier.

The following examples illustrate certain embodiments of this invention and are not intended to limit the scope of this invention.

Embodiments herein are further described by the following examples and detailed protocols. However, the examples are only intended to illustrate the embodiments and should not be construed as limiting the scope of the specification. The contents of all references and published patents and patent applications cited throughout this application are hereby incorporated by reference.

[Example 1] P. Acnes bacteriophage PHIT-101 is a P. Selectively and efficiently sterilize Acnes P. Acnes bacteriophages are highly genetically similar and are described in P. It has been shown to exhibit a wide range for multiple strains of Acnes. Lead bacteriophage (PHIT-101) was used for experiments. PHIT-101 is a P. It is the only lytic phage that killed all strains of Acnes (data not shown). PHIT-101 has the sequence of SEQ ID NO:1. To introduce the efficacy and specificity of this phage, a plate assay was performed as follows. P. Acnes KPA171202 and P.P. Granulosum (a closely related but benign skin bacterium) was plated on separate BHI agar plates. A sterile cotton pad was placed on each plate. Sterile cotton pads were dipped in minocycline, an antibiotic commonly used to treat acne, or in a phage solution with a titer of 2×10 ⁷ pfu/mL. After anaerobically incubating the plates at 37° C. for 72 hours, the minocycline pads indiscriminately killed the bacteria and caused P. Acnes and Symbiosis P. Both granulosum showed a germicidal zone (Fig. 1). In contrast, the PHIT-101 pad has a P. It sterilizes only Acnes and gives a beneficial P. It did not hinder the growth of granulosum.

Further evidence of PHIT-101's ability to selectively sterilize was obtained in a synthetic skin microbiome assay. The three skin bacteria that make up 60-80% of the bacterial flora in skin pores, P. Acnes, P. Granulosum and P. A synthetic skin microbiome containing avidum was devised [Science (2009) 324: 1190-1192]. This synthetic skin microbiome was grown anaerobically in the presence or absence of PHIT-101 (final concentration 5×10 ⁵ pfu/mL). After 48 hours incubation at 37° C., cells were pelleted and washed and the relative proportions of the three strains were determined using 16S amplicon next generation sequencing (NGS) on an Illumina MiSeq. According to the result shown in FIG. Acnes can be almost completely sterilized, and symbiotic P. Granulosum and P. It has been shown to have no negative impact on the growth of Abidum.

P. Screening for biofilm degrading enzymes (BDEs) that destroy the Acnes biofilm. Several recent reports (Exp Dermatol (2014) 23:687, Br J Dermatol (2015) 172:13), P. It has been established that Acnes produces significant amounts of biofilm in skin pores, which impedes antibiotic penetration and results in poor therapeutic outcome. To verify this, P. Biofilm production of several strains of Acnes was quantified. FIG. 3 shows that adherent cultures of multiple strains isolated from a single subject's flora produce significantly different levels of biofilm under similar conditions. Therefore, a previous proof of concept using floating cells was described in P. Acnes did not reflect the true condition of growth on the skin.

Without being bound to any scientific theory, the inventors have found that biofilms adhere to P. We hypothesized that it could be a significant barrier to phage killing of Acnes cells. This hypothesis is based on floating P. Unlike Acnes (99% sterilization, Fig. 2), PHIT-101 is a P. This was confirmed in the cell viability assay (Figure 5) which showed that only about 50% of Acnes cells could be killed. To determine if biofilm degradation improves phage killing, P. Several enzymes were screened to find BDE specific to acnes. The screen included three classes of enzymes capable of degrading the types of substances that might be found in biofilms: DNA, polysaccharides and proteins. Figure 4 shows that in the screen, deoxyribonuclease has moderate activity and the best biofilm degradation rate is with dispersin, which is a glycoside hydrolase from proteases and Aggregatibacter actinomycetemcomitans. Indicates that it has been found.

In the selection of BDE paired with phage, dispersin was selected for two reasons. First, as a glycoside hydrolase, dispersin was unable to attack the protein membrane of the phage itself, thereby avoiding possible degradation of the phage. Secondly, P. Acnes forms a robust biofilm with Staphylococcus aureus [Anaerobe (2016) 40:63-67] and dispascin is active against biofilms from both organisms. By adding dispersin, fixed P. It was determined whether the efficiency of phage killing in Acnes increased. FIG. 5 shows that the killing of PHIT-101 was enhanced in the presence of dispersin, restoring the killing efficiency of about 99% of phage.

Example 2 Probiotic Bacteria Genotyping Characterization of Probiotic Strains P. Strains of Acnes are based on point mutations in the 16S rDNA sequence leading to pathogenicity and phylogenetic classification into probiotic strain types and the absence of linear plasmids found in pathogenic strains carrying virulence factors. Was characterized by. Using the 16S specific primer, each P. The complete 16S rDNA sequence of the Acnes strain was amplified and Sanger sequenced. It was confirmed that the probiotic strain has ProI and ProII ribosequences (RS). The ProI strain has the T838C and C1322T mutations as compared to the 16S rDNA sequence of the KPA171202 reference strain (NIH accession number NC — 006085.1). The ProII strain has the C986T and T992C mutations compared to the KPA171202 sequence. In addition, specific primer pairs were used to determine the presence or absence of linear plasmids within each strain. The probiotic strain was confirmed to lack this plasmid, which carries an extrachromosomal lipase as well as a Tad (adhesion) virulence factor.

In an embodiment, the probiotic strain is characterized primarily by its 16S sequence, eg, SEQ ID NO:3 and SEQ ID NO:4. In an embodiment, probiotic strains can be genotyped by lacking plasmids carrying virulence factors such as extrachromosomal lipases and the Tad locus.

A cohort of probiotic strains was further characterized for their immunogenic potential. The lead probiotic candidate was based on two factors: low lipase production and reduced adherence to epithelial cells. Phenotypic confirmation of these features was important in selecting probiotic lead candidates.

Probiotic P. Testing the immunogenic potential of the Acnes strain: Lipase activity Lipase plays an important role in the pathogenesis of acne by hydrolyzing sebum triglycerides and releasing stimulating free fatty acids into the pilosebaceous follicles. Plays. Lipase is a strong chemotactic and proinflammatory antigen. Therefore, lipases are of great interest as pharmacological targets for anti-acne drugs. In an embodiment, the overall strategy is a pathogenic P. coli that secretes high levels of lipase. Acnes is a low-secretory probiotic P. To replace with Acnes. To quantify the lipase expression phenotype for each of our panel strains, the probiotic P. The lipase production of the Acnes strain was determined using the fluorescent lipase activity assay for pathogenic P. Compared to Acnes strain.

One of the most interesting findings was that each strain secreted different amounts of lipase when grown in suspension versus adherent culture. This is P. Previously reported in the Acnes strain [Res Microbiol, (2007) 158:386-392]. Furthermore, the data showed that when these strains were grown in liquid culture, there was no significant difference between the pathogenicity and the lipase production of the probiotic strains. However, interesting changes were seen when these strains were grown under biofilm conditions. Variability of production between strains could still be observed, but some probiotic strains had significantly less lipase activity than pathogenic strains (Figure 11). Interestingly, not all strains in the probiotic cohort had low lipase activity. For example, the lipase production of strains Pr-1 and Pr-5 was above the threshold of probiotic strains and did not progress any further. Therefore, sticking P. By quantifying lipase production in Acnes cells, it was possible to screen among probiotic strains and select lead candidates with the most consistently low levels of lipase activity.

Thus, pathogenic and probiotic strains secreted similar amounts of lipase in suspension cultures, but in adherent cultures probiotic strains secreted much less lipase than pathogenic strains. FIG. 8 shows that the top probiotic candidates had a lower lipase profile compared to the pathogenic strain.

Probiotic P. Testing the immunogenic potential of the Acnes strain: cell adhesion The available pathogenic strains were confirmed to have a cohesive (tad) locus that plays a role in the pathogenicity of other mammalian pathogens [J. Bacteriol (2000) 182:6169-6176; Nat Rev Microbiol (2007) 5:363-375; PNAS (2003) 100:7295-7300]. Greater adhesion to host cells can enhance pathogenicity or induce an inflammatory host response. Adhesion to epithelial cells was expected to be less robust because probiotic strains were previously genotyped to not contain the tad locus. To assess whether there was a clear difference in adhesion, the adhesion of pathogenic and probiotic strains to A-431 skin epithelial cells was compared. FIG. 9 shows that the top three probiotic candidates did not adhere as strongly to epithelial cells as the pathogenic strains. Interestingly, a subtle but obvious difference in cell adhesion is P. Rediscovered among different strain families of Acnes. As described above, P. The Acnes strain showed slightly higher cell adhesion (Pr-2 in FIG. 9) and the ProII strain showed less robust cell adhesion (Pr-B, Pr-C in FIG. 9).

Pathogenicity in mouse ear inflammation model and probiotic P. Comparing Acnes, showing less lipase production, less adherence to epithelial cells and demonstrating the lower immunogenic potential of probiotic strains, the inflammatory response of these strains was As a well-established model, P. in the context of acne has been used. The test was performed with the mouse ear inflammation model that has been used to assess the proinflammatory properties of Acnes. The pro-inflammatory properties of pathogenic and probiotic strains were compared in the following studies. 10 ¹⁰ cfu strains were injected into the ears of CBA/J mice. A cohort of 5 mice was assigned to each strain. Five days later, the ears were excised and examined for inflammation. The levels of several inflammatory cytokines (IL-1β, IL-6, IL-17, TNFα) were measured and tissue sections examined by histological examination. FIG. 10 shows that the pathogenic strain had significantly higher levels of IL-1β, IL-6, IL-17 and TNFα compared to the probiotic strain.

Acute Skin Safety and Toxicity of Probiotic Strains in a Minipig Skin Model Probiotic strains were tested for skin irritation using a minipig model. Pigs are one of the major animals used in translational research, and pig skin is physiologically, anatomically, biochemically and immunologically similar to human skin. Minipigs, in particular, are commonly used to model human skin diseases and conditions such as acne [Vet Pathol (2012) 49:344-356]. The probiotic strain was applied at two doses, 10 ⁸ cfu and 10 ⁹ cfu, to delimited skin areas of the skin of three separate minipigs. Animals were observed daily for clinical signs and skin at the dosing site was scored using the Draize scoring system before dosing, 0.5, 1, 4, 8 and 24 hours after dose administration. There was no erythema or edema associated with the lead probiotic strain during the entire period (Table 1) and a Draize score of 0 was observed throughout. This demonstrates the safety of our probiotic strain against acute exposure in an animal skin model.

Table 1: Acute skin safety/toxicity in a minipig skin model indicates a good safety profile for probiotic strains. The probiotic strain was applied at normal (10 ⁸ cfu) and acute (10 ⁹ cfu) doses to delimited skin areas in 3 male minipigs and monitored for 24 hours after application. Erythema and edema were quantified using the Draize scoring method. The Draize score provides the relative severity of erythema and edema. A Draize score of 0 indicates no erythema and edema, but was observed in all skin areas throughout the monitoring period.

Example 3 Stability of Bacteriophage in Compositions with Anti-acne Compounds To determine if phages are stable in combination with salicylic acid or benzoyl peroxide (BPO), phages are tested. Co-incubation with these agents at low and high concentrations. The range of concentrations was determined by the approved concentrations of these agents specified in the US Food and Drug Administration (FDA) Acne Monograph for OTC Drugs. For salicylic acid this is 0.5% to 2% (w/v) and for BPO the range is 2.5% to 10% (w/v). Phage buffer was added to these agents and their stability at 4°C was tested for 60-90 days. Figure 15 shows that the phage are stable in the presence of both low and high doses of salicylic acid. In contrast, Figure 16 shows that benzoyl peroxide destabilizes the phage and the observed rate of decrease in phage viability is steeper at higher concentrations of BPO.

Example 4 (Virtual Example) Treatment with a Combination of Bacteriophage and Salicylic Acid To determine the efficacy comparison of this treatment with placebo, Propionibacterium acnes bacteriophage alone and salicylic acid alone, propionibacteria A double-blind, placebo-controlled study of a composition comprising Um acnes bacteriophage and salicylic acid is conducted. Salicylic acid at concentrations of 0.5% and 2% (w/v) is administered with and without Propionibacterium acnes bacteriophage. In all conditions, including Propionibacterium acnes bacteriophage, the phage is present at a dose of 10 ⁹ pfu (plaque forming units) per dose. Ten subjects with relatively severe acne will be treated by the following groups:
(I) Placebo (no active drug)
(Ii) 0.5% salicylic acid as the only active agent (iii) 2% salicylic acid as the only active agent (iv) Propionibacterium acnes bacteriophage (v) 0.5% as the only active agent Combination of salicylic acid and Propionibacterium acnes bacteriophage (in a single composition)
(Vi) A combination of 2% salicylic acid and Propionibacterium acnes bacteriophage (in a single composition).

The combination of Propionibacterium acnes bacteriophage and salicylic acid achieves an additive or more synergistic effect in the treatment of acne (the combined effect of bacteriophage and salicylic acid is Greater than the sum of the effects of bacteriophage and salicylic acid. Efficacy of treatment is measured using lesion count and IGA (Investigator Overall Assessment) score.

Example 5 Virtual Example) Treatment with a Combination of Bacteriophage and Sulfur To determine the efficacy comparison of this treatment with placebo, Propionibacterium acnes bacteriophage alone and sulfur alone, Propionibacterium Perform a double-blind, placebo-controlled study of compositions containing acnes bacteriophage and sulfur. Sulfur at concentrations of 3% and 10% (w/v) is administered with and without Propionibacterium acnes bacteriophage. In all conditions, including Propionibacterium acnes bacteriophage, the phage is present at a dose of 10 ⁹ pfu per dose. Ten subjects with relatively severe acne will be treated by the following groups:
(I) Placebo (no active drug)
(Ii) 3% sulfur as the only active agent (iii) 10% sulfur as the only active agent (iv) Propionibacterium acnes bacteriophage (v) 3% sulfur and propioni as the only active agent Combination of bacterium acnes bacteriophage (in a single composition)
(Vi) 10% sulfur in combination with Propionibacterium acnes bacteriophage (in a single composition)

The combination of Propionibacterium acnes bacteriophage and sulfur achieves an additive or more synergistic effect in treating acne (the combined effect of bacteriophage and sulfur is that each drug is used separately). Greater than the sum of the effects of bacteriophage and sulfur if they are. Efficacy of treatment is measured using lesion count and IGA (Investigator Overall Assessment) score.

Example 6 (Virtual Example) Treatment with a Combination of Bacteriophage and Benzoyl Peroxide To determine the efficacy comparison of this treatment with placebo, Propionibacterium acnes bacteriophage alone and BPO alone. A double-blind, placebo-controlled study of a composition containing Pionibacterium acnes bacteriophage and BPO is conducted. BPO at concentrations of 2.5% and 10% (w/v) is administered with and without Propionibacterium acnes bacteriophage. In all conditions, including Propionibacterium acnes bacteriophage, the phage is present at a dose of 10 ⁹ pfu per dose. Ten subjects with relatively severe acne will be treated by the following groups:
(I) Placebo (no active drug)
(Ii) 2.5% BPO as the only active agent
(Iii) 10% BPO as the only active drug
(Iv) Propionibacterium acnes bacteriophage as the only active agent (v) 2.5% BPO in combination with Propionibacterium acnes bacteriophage (in a single composition)
(Vi) 10% BPO in combination with Propionibacterium acnes bacteriophage (in a single composition)

The combination of Propionibacterium acnes bacteriophage and BPO achieves an additive or more synergistic effect in treating acne (the combined effect of bacteriophage and BPO is that each drug is used separately). Greater than the sum of the effects of bacteriophage and BPO when given). Efficacy of treatment is measured using lesion count and IGA (Investigator Overall Assessment) score.

Example 7 (Virtual Example) Assay using a combination of bacteriophage and benzoyl peroxide. Suspension and adhesion. In vitro tests have been performed to determine the efficacy of (i) BPO; (ii) Propionibacterium acnes bacteriophage; or (iii) Propionibacterium acnes bacteriophage + BPO in killing Acnes bacteria. It

The combination of Propionibacterium acnes bacteriophage and BPO was associated with sessile pathogenic P. Achieves an additive effect or more in killing Acnes bacteria, that is, a synergistic effect. large.). The keratolytic action of BPO (similar to salicylic acid and retinoids) is due to the fact that phage penetrates the skin pores and is deep inside the pores. Help approach Acnes.

Claims (55)

A composition comprising at least one Propionibacterium acnes bacteriophage, at least one anti-acne compound and a pharmaceutically acceptable carrier. The composition of claim 1, which is free of probiotic bacteria. The composition is P. The composition of claim 1, further comprising an Acnes biofilm degrading enzyme. The composition of claim 1, wherein the at least one anti-acne compound is salicylic acid. The composition of claim 4, wherein salicylic acid is present at a concentration of 0.5% to 2% (weight/volume). 6. The composition of claim 5, wherein salicylic acid is present at a concentration of less than 0.5% but greater than about 0.1% (weight/volume). The composition of claim 1, wherein the at least one anti-acne compound is sulfur. The composition of claim 7, wherein sulfur is present in a concentration of 3% to 10% (weight/volume). Sulfur is less than 3% but is present in a concentration of greater than about 0.1%, 0.5%, 1%, 1.5%, 2% or 2.5% (weight/volume), The composition according to claim 7. The composition of claim 1, wherein the at least one anti-acne compound is resorcinol and sulfur. 11. The composition of claim 10, wherein resorcinol is present in a concentration of 2% and sulfur is present in a concentration of 3% to 8% (weight/volume). The composition of claim 1, wherein the at least one anti-acne compound comprises resorcinol monoacetate and sulfur. 13. The composition of claim 12, wherein resorcinol monoacetate is present in a concentration of 3% and sulfur is present in a concentration of 3% to 8% (weight/volume). The composition according to claim 1, wherein the anti-acne compound is an antibiotic, a retinoid or an alpha hydroxy acid. The composition of claim 1, wherein the Propionibacterium acnes bacteriophage is a naturally occurring Propionibacterium acnes bacteriophage. P. Acnes bacteriophage is a lytic P. The composition of claim 1 which is an acnes bacteriophage. P. The composition of claim 1, wherein the Acnes bacteriophage comprises a linear double stranded DNA genome. P. The composition of claim 1, wherein the Acnes bacteriophage is within the family of Bacteriophage Siphoviridae. P. The Acnes bacteriophage genome comprises a nucleotide sequence that is at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to the nucleotide sequence of SEQ ID NO:1. The composition according to. The enzyme is P. The composition according to claim 3, which is an Acnes biofilm degrading enzyme. 21. The composition of claim 20, wherein the enzyme is a glycosidase, protease, deoxyribonuclease or restriction endonuclease. 21. The composition of claim 20, wherein the enzyme is glycosidase. 23. The composition of claim 22, wherein the glycosidase is a glycoside hydrolase. 24. The composition of claim 23, wherein the enzyme catalyzes the hydrolysis of a linear polymer of N-acetyl-D-glucosamine. 25. The composition of claim 24, wherein the enzyme is β-hexosaminidase. 26. The composition of claim 25, wherein the enzyme hydrolyzes the β-1,6-glycosidic bond of the acetylglucosamine polymer. 21. The composition of claim 20, wherein the enzyme is deoxyribonuclease I, restriction endonuclease, papain, bromelain, trypsin, proteinase K, subtilisin, seratiopeptidase, dispasin, alginate lyase, amylase or cellulase. 21. The composition of claim 20, wherein the enzyme is Dispersin B. 21. The composition of claim 20, wherein the enzyme is a protease and the protease is proteinase K or subtilisin. The composition of claim 1, further comprising an anti-aging enzyme. 31. The composition of claim 30, wherein the anti-aging enzyme is superoxide dismutase or peroxidase. The composition of claim 1, further comprising probiotic bacteria. The probiotic bacteria are probiotic P. 33. The composition of claim 32, which is a bacterial species (P. sp.), Staphylococcus species and/or Corynebacterium spp. 33. The composition of claim 32, wherein the probiotic bacterium is a bacterium within the Betaproteobacteria class. The probiotic bacteria are probiotic P. 34. The composition of claim 33, which is an Acnes bacterium. P. Acnes bacteria
(A) contains the 16S rDNA sequence with the T992C mutation as compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2;
(B) contains the 16S rDNA sequence with the T838C mutation as compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2;
(C) contains a 16S rDNA sequence with the C1322T mutation as compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2;
(D) contains the 16S rDNA sequence with the C986T mutation as compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2;
(E) comprises a 16S rDNA sequence that is identical to the sequence of SEQ ID NO:3;
(F) comprises a 16S rDNA sequence that is identical to the sequence of SEQ ID NO:4;
(G) does not contain a linear plasmid;
(H) does not contain a plasmid containing a virulence factor; and/or (i) does not contain a plasmid encoding an extrachromosomal lipase and/or an adherent virulence factor,
36. The composition of claim 35. P. Acnes bacteria
(A) Pathogenic P. cerevisiae when grown in suspension culture. Produces less than about 20% of the level of lipase produced by the Acnes strain;
(B) Pathogenic P. cerevisiae when grown in adherent culture. Produces less than about 10% of the level of lipase produced by the Acnes strain;
(C) Adhesion to epithelial cells causes pathogenic P. At least 50% less than the Acnes strain, and/or (d) pathogenic P. Less inflammatory than Acnes strain,
36. The composition of claim 35. 36. The composition of claim 35, further comprising at least one additional probiotic bacterium. At least one additional probiotic bacterium is Propionibacterium granulosum.
39. The composition of claim 38, which comprises and/or Propionibacterium avidum. Pathogenic P. Acnes strains,
(A) contains the 16S rDNA sequence with the G1058C mutation as compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2;
(B) containing the 16S rDNA sequence with the G1058C and A1201C mutations compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2;
(C) contains the 16S rDNA sequence with the G529A mutation as compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2;
(D) contains the 16S rDNA sequence with the G1004A and T1007C mutations compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2;
(E) contains the 16S rDNA sequence with the G1268A mutation as compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2;
(F) contains the 16S rDNA sequence with the T554C and G1058C mutations compared to the KPA171202 reference strain 16S rDNA sequence shown as SEQ ID NO:2;
(G) contains a 16S rDNA sequence that is identical to the sequence of SEQ ID NO:5;
(H) comprises a 16S rDNA sequence that is identical to the sequence of SEQ ID NO:6;
(I) comprises a 16S rDNA sequence that is identical to the sequence of SEQ ID NO:7;
(J) comprises a 16S rDNA sequence which is identical to the sequence of SEQ ID NO:8;
(K) comprising a 16S rDNA sequence identical to the sequence of SEQ ID NO: 9; and/or (l) comprising a 16S rDNA sequence identical to the sequence of SEQ ID NO: 10,
38. The composition of claim 37. At least one additional P. The composition of claim 1, further comprising an acnes bacteriophage. The composition of claim 1, wherein the pharmaceutically acceptable carrier comprises an emulsion. 43. The composition of claim 42, wherein the emulsion is an oil-in-water emulsion or a water-in-oil emulsion. The composition according to claim 1, which is in the form of a cream, lotion, suspension or aqueous solution. A combination comprising at least one Propionibacterium acnes bacteriophage and at least one anti-acne compound, wherein each of the at least one Propionibacterium acnes bacteriophage and at least one anti-acne compound is a pharmaceutical A combination in the composition further comprising a pharmaceutically acceptable carrier. At least one P. 46. The combination of claim 45, wherein the acnes bacteriophage and the at least one anti-acne compound are in separate compositions. 47. The combination of claim 46, wherein the at least one anti-acne compound is benzoyl peroxide. 48. The combination of claim 47, wherein the benzoyl peroxide is present at a concentration of 2.5%-10% (weight/volume). Benzoyl peroxide is less than 2.5% but is present in a concentration of greater than about 0.1%, 0.5%, 1%, 1.5% or 2% (weight/volume). 47. The combination according to 47. A method of treating acne in a subject in need thereof, comprising administering to the subject an effective amount of the composition of claim 1. 51. The method of claim 50, wherein the composition is administered topically. 46. A method of treating acne in a subject in need thereof, comprising administering to the subject an effective amount of the combination of claim 45. A composition comprising Propionibacterium acnes bacteriophage and an enzyme. A combination comprising Propionibacterium acnes bacteriophage and an enzyme. A composition consisting essentially of at least one Propionibacterium acnes bacteriophage, at least one anti-acne compound and a pharmaceutically acceptable carrier.

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